http://2008.igem.org/wiki/index.php?title=Special:Contributions/Joyy&feed=atom&limit=50&target=Joyy&year=&month=2008.igem.org - User contributions [en]2024-03-29T09:20:36ZFrom 2008.igem.orgMediaWiki 1.16.5http://2008.igem.org/Team:Harvard/ProjectTeam:Harvard/Project2008-10-30T05:03:13Z<p>Joyy: </p>
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==BACTRICITY: Bacterial Biosensors with Electrical Output==<br />
<br />
<br />
The metabolically versatile bacterium ''Shewanella oneidensis'' adapts to anaerobic environments by transporting electrons to its exterior, reducing a variety of environmental substrates. When grown anaerobically and provided with lactate as a carbon source, ''S. oneidensis'' transfers electrons to an electrode of a microbial fuel cell. We sought to engineer ''S. oneidensis'' to report variations in environmental conditions through changes in current production. A previous study has shown that ''S. oneidensis'' mutants deficient in the mtrB gene produce less current than the wildtype strain, and that current production in these mutants can be restored by the addition of exogenous mtrB. We attempted to control current production in mtrB knockouts by introducing mtrB on lactose, tetracycline, and heat inducible systems. These novel biosensors integrate directly with electrical circuits, paving the way for the development of automated, biological measurement and reporter systems.<br />
<br />
==Experimental overview==<br />
<br />
We attempted to develop three inducible systems for electrical current production in ''S. oneidensis''. The first is a chemically inducible system, where LacI or TetR repression of the current-production gene expression in mtrB knock-out (KO) ''S. oneidensis'' can be alleviated by the addition of IPTG or anhydrotetracycline, respectively. Our second approach uses temperature-senstive cI which would allow for an increase in electrical output in response to heat. The third is a light-inducible system based on the 2005 UT Austin biological camera.<br />
<br />
Using the LacI system, we attempted to interact with engineered ''S. oneidensis'' in multiple anaerobic microbial fuel cells which we built, measuring the effects of IPTG on current production in wild type and mtrB KO ''S. oneidensis'' containing plasmids expressing inducible or constitutive mtrB.<br />
<br />
==Results==<br />
<br />
===Wildtype vs. mtrB deficient ''S. oneidensis''===<br />
<br />
''Overview: By genetically manipulating S. oneidensis, it has been found that noticeable differences of current production can be detected. We wish to control current production by introducing inducible systems in these mutant strains of S. oneidensis. These differences of current production can be detected by a computer, allowing biological systems and electrical devices to be integrated.''<br />
<br />
The basis of our project lies in the discovery that by knocking out the mtrB gene, a current production gene of S. oneidensis, noticeable differences of current production can be detected. Therefore, this allows the current production of S. oneidensis to act more than just an on/off switch.<br />
<br />
The first step of our project was then to test this hypothesis and also to understand the current production behavior of wildtype S. oneidensis and the mutant S. oneidensis (annotated as mtrB). Therefore, we ran a test with microbial fuel cells, one strain in each cell, and allowed the current production to be measured for approximately one day. In literature, mtrB produces approximately 20 to 25 percent of the current or wildtype strains.<br />
<br />
<br />
[[Image: wt_mtrB.jpg|700px]]<br />
<br />
<br />
As shown in the graph above, the results of our experiment matched those in literature. We also observed that wildtype and mtrB strains have very different current production behavior, with wildtype producing current immediately after lactate injection, and mtrB producing current much more gradually. This difference in behavior can allow for more accurate differentiation of current production between wildtype and mtrB strains when integrated with electrical devices. We wish then build on these results and build inducible systems that will allow us to control the current production.<br />
<br />
===Lac-inducible Strains===<br />
<br />
''Overview: We genetically engineered mtrB knock-out S. oneidensis MR-1 by introducing the mtrB gene on a lactose-inducible system. Specifically, we tested engineered mtrB knock-out S. oneidensis MR-1 with high lacQPI at the p15A origin. Our results show the possibility that we successfully complemented the mtrB knock-out as high levels of current were detected in one such strain.''<br />
<br />
Our end goal was to develop inducible systems for electrical current production in S. oneidensis MR-1. In our microbial fuel cells, we were able to test a lac-inducible system where LacI repression of the current-production gene expression in mtrB knock-out S. oneidensis MR-1 would be alleviated by the addition of IPTG. That is, the addition if IPTG to such a system would induce current production as the bacteria would begin breaking down lactate.<br />
<br />
In this experiment, we tested the following combinations:<br />
<br />
1. wt S. oneidensis MR-1 + lactate <br />
<br />
2. mtrB knock-out S. oneidensis MR-1 with lactate <br />
<br />
3. mtrB (with high Lac promoter) with lactate and IPTG<br />
<br />
4. mtrB (with high Lac promoter) with lactate<br />
<br />
We expected combination 1 to be highest, peaking between 100 to 200 microAmps. Combination 2 would set the baseline for the mtrB knock-out strain, usually peaking at 20 to 25 percent of wt S. oneidensis MR-1’s level. If the introduction of mtrB on a lactose-inducible system was successful, then combination 3 would also be high, roughly around the level of wt S. oneidensis MR-1. Combination 4, however, would be expected to peak around the mtrB knock-out strain’s current level as it did not receive IPTG and would not be induced.<br />
<br />
[[ Image: Lacinducible.jpg | 700px]]<br />
<br />
As expected, wt S. oneidensis MR-1 and the mtrB knock-out strain reached their expected levels. Interestingly, combination 4, but not combination 3, reached current levels around 200 microAmps, even greater than wt S. oneidensis MR-1’s level. That is, the engineered mtrB knock-out S. oneidensis MR-1 that did not receive IPTG resulted in elevated current production. The temporal dynamics of the curve was what we would expect from IPTG induction. That is, a delay would be observed as IPTG induction took place turning on the promoter relative to the non-inducible wt S. oneidensis MR-1. <br />
<br />
A possible, and exciting, explanation for this observance may stem from the fact that the repressor used in our engineered strain had an LVA tag which marks it for degradation. As the experiment progressed, the repressor proteins in combination 4 may have degraded, which allowed for lactate breakdown and thus current production. The implication is that if the current production observed is due to the degradation of the repressor protein, then we successfully complemented the mtrB knock-out. We must note that if this were the case, then we should have seen an increase in all strains of the engineered mtrB knock-out if the repressor protein was being degraded. It may be that we would have observed this increase if we had run the experiment for a longer period. Future experiments may shed more light on this observance.<br />
<br />
===Co-Culture Experiment===<br />
<br />
''Overview: One possible system for achieving inducible current could be to couple a inducible-lacZ system in E. coli to current production by wildtype S. oneidensis MR1 through the conversion of lactose to lactate. This leaves open the possibility of using a pre-existing E. coli lacZ reporter or creating our own.<br />
''<br />
<br />
In addition to genetically engineering S. oneidensis MR-1 to respond to chemicals and heat, we also sought to take advantage of its natural metabolic pathway of breaking down lactate to produce current. From our wildtype versus mtrB knockout experiments, we found that feeding S. oneidensis MR-1 lactate led to significant current production almost instantly. In wildtype S. oneidensis MR-1, the current production would increase and stay elevated for a period of twelve hours. This result occurred consistently, thus we sought to use controlled lactate release as a way to control current output. <br />
<br />
We also knew that wildtype E. coli takes lactose and breaks down lactate. Thus, we hypothesized that it might be possible to couple E. coli’s lactate production with S. oneidensis MR-1’s lactate breakdown to produce current. In addition, a great deal of genetic engineering has already been done on E. coli and the up-regulation or down-regulation of its Lac operon. This means that instead of genetically modifying S. oneidensis MR-1 to respond to chemicals or heat, we can instead take advantage of the vast library of E. coli genetics and couple genetically engineered E. coli with wildtype S. oneidensis MR-1. <br />
<br />
In this experiment, we tested different combinations of wildtype E. coli MG1655, S. oneidensis MR-1, and Lac-operon knockout E. coli MC4100 as follows:<br />
<br />
1. wt E. coli (MG1655) + wt S. oneidensis (MR1) + lactose <br />
<br />
2. wt E. coli (MG1655) + wt S. oneidensis (MR1) + lactate pos. control<br />
<br />
3. Lac-operon knockout E. coli (MC4100) + wt S. oneidensis (MR1) + lactose<br />
<br />
4. Lac-operon knockout E. coli (MC4100) + wt S. oneidensis (MR1) + lactate pos. control<br />
<br />
5. wt S. oneidensis MR-1 + lactose neg. control<br />
<br />
6. wt E. coli (MG1655) + lactose neg. control<br />
<br />
7. Lac-operon knockout E. coli (MC4100) + lactose neg. control<br />
<br />
Based on previous experiments, we would expect current production in combinations 2 and 4 as they both have wildtype S. oneidensis MR-1 and receive lactate. In addition, however, we would expect combination 1 to also produce current. As described above, wt E. coli would break down lactose into lactate, and S. oneidensis MR-1 would break down lactate to produce current.<br />
<br />
[[ Image: picture 5.png | 750px ]]<br />
<br />
From the data above, we found that combination 1 did indeed produce current with a delay relative to the positive control. The delay can be attributed to the time it takes for E. coli to break down lactose into lactate, thus adding an extra step in the carbon source to current production pathway compared to our positive controls. These results are exciting in that they show a possibility of taking advantage of this cooperative effort to achieve inducible current.<br />
<br />
==Future directions==<br />
<br />
Our work with creating a system of inducible electrical output in ''S. oneidensis'' has laid the foundations for many different exciting avenues of further inquiry which look to take advantage of a bacteria-computer interface that combines the amazing sensitivity and adaptability of bacteria with the speed and analytical abilities of electricity and computers.<br />
<br />
Using the same principles underlying the lac system, the [https://2006.igem.org/University_of_Edinburgh_2006 arsenic biosensor] developed by the University of Edinburgh iGEM 2006 team could be introduced into ''S. oneidensis'', allowing for the coupling of arsenic sensing to an electrical output, a form of a data which is easier to automate and transmit. This could be further extended to other chemical sensing systems, such as the [https://2007.igem.org/Brown lead sensor] created by the Brown iGEM 2007 team and the [https://2007.igem.org/MIT mercury sensor] made by the MIT iGEM 2007 team, resulting ultimately in an array of different strains of ''S. oneidensis'' which all respond to the presence of different chemicals with an electrical output that can be monitored by a computer. This could theoretically allow for the remote sensing and analysis of the chemical composition of an environment over time in a cost-effective manner, making it a tool with powerful public health applications, such as monitoring water quality.<br />
<br />
Another interesting direction would be the linking of the [https://2006.igem.org/UT_Austin_2005 light-sensing system] developed by the UT Austin iGEM team with electrical output in ''S. oneidensis''. In response to variations in light, the amount of electricity produced by ''S. oneidensis'' would change. This would allow for the intriguing possibility of not only ''S. oneidensis'' conveying information to the computer, but also the computer responding to the ''S. oneidensis''. A simple example would be that in response to a chemical input, ''S. oneidensis'' may increase its electrical output. Sensing this increase, the computer could turn on or off a light directed at the ''S. oneidensis'', modifying ''S. oneidensis'''s output, creating interesting feedback loops. This could ultimately be developed into more complex communications systems between bacteria and computers. We tried constructing this system over summer, but as the process requires making an EnvZ knockout strain of ''S. oneidensis'', we could not finish it. We did, however, make a few parts to facilitate future attempts.<br />
<br />
The possibilities are further broadened by our observations of co-cultures of ''E. coli'' and ''S. oneidensis''. Either of the systems described above could be pursued through an alternative alternative strategy of co-cultures. For instance, an array of ''E. coli'' which respond to different chemicals by breaking down lactose into lactate could be cultured with ''S. oneidensis''. In response to an increase in lactate, ''S. oneidensis'' would begin to produce higher levels of electricity. Co-cultures could also allow for more complex bacteria-computer interactions. This strategy could enable the coupling of almost any ''E. coli'' ability to electrical output.<br />
<br />
These future directions in which our research can be taken demonstrate some of the exciting possibilities of BACTRICITY!<br />
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|}</div>Joyyhttp://2008.igem.org/Team:Harvard/HardwareTeam:Harvard/Hardware2008-10-30T04:12:15Z<p>Joyy: /* Software */</p>
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=Fuel Cell Development=<br />
==Motivation==<br />
<br />
The broad goal of our project was to engineer ''S. Oneidensis'' to produce a detectable change in electric current in response to some environmental stimulus. In order to observe such a reaction, our first task was to design an environment capable of housing bacteria and measuring current production.<br />
<br />
==Solution - Microbial Fuel Cells==<br />
<br />
===Background===<br />
<br />
To measure the electric current production in Shewanella oneidensis MR-1, we constructed microbial fuel cells (MFCs) to harvest the electricity produced. Traditional fuel cells convert chemical energy to electrical energy. MFCs are unique in that the fuel source used is microbially degradable organic matter (Lovley 2006). Such organic matter is a form of renewable energy and can come from materials in wastewater, sediments, or agricultural wastes (Cho et al. 2007). Compared to hydrogen- and methanol-driven fuel cells, MFCs are particular attractive due to their ease of operation and wide range of fuel sources available. They do not require expensive catalysts, high operating temperatures, nor explosive or toxic fuels (Lovley 2006). Indeed, we found the construction and operation of our microbial fuel cells to be relatively straightforward. <br />
<br />
MFCs operate using principles similar to normal fuel cells. MFCs require an anode and a cathode separated by a semi-permeable membrane (permeable to protons) (Lovley 2006). The anode compartment is generally anaerobic and is the site of oxidation of organic matter (Lovley 2006); the cathode transfers electrons to the electron acceptor (Lovley 2006). MFCs take advantage of the natural metabolism of microorganisms that break down organic matter by intercepting the electrons that would be donated to naturally occurring electron acceptors. Instead, these electrons are transferred from the anode to the cathode, usually along metal wires to which current readings can be taken. <br />
<br />
A figure of a microbial fuel cell is shown below (NB: this is not what we used in our experiments). <br />
<br />
[[ Image: Snapshot1.png | 350px ]]<br />
<br />
On the anode side, glucose is being metabolized by the microorganism to yield electrons which are transferred to the cathode side where oxygen is reduced to water (Lovley 2006).<br />
<br />
===Context===<br />
<br />
Shewanella oneidensis MR-1 is a suitable microorganism for a MFC. It is known to break down lactate (Bretschger et al. 2007), and in anaerobic conditions, S. oneidensis MR-1 produces nanowires that shuttle electrons to electron acceptors as its aerobic electron acceptor, oxygen, is unavailable (Gorby et al. 2006). Taking advantage of this, we designed our microbial fuel cell with an anaerobic anode where completion of the lactate reduction pathway must be completed by transferring electrons to the cathode side at which reduction of oxygen to water occurs. The anode chamber, besides housing our bacteria, was also the site of chemical injections. We provided S. oneidensis MR-1 with lactate as its sole carbon source and sought to induce the mtrB gene, thus “turning on” current production, with chemicals such as lactose and tetracycline. In addition, our heat tests were directed towards the anode chamber as well. Thus, the detection of electricity production of S. oneidensis MR-1 using MFCs serves as an innovative way to detect gene expression, combining both traditional molecular biology with renewable energy engineering. <br />
<br />
<br />
[[Image:Chambers.jpg|600px]]<br />
<br />
==Design Goal==<br />
<br />
===Functional description===<br />
<br />
The final product is a complete system capable of introducing separate strains of bacteria to multiple different environments while simultaneously measuring and recording current readings from each. The experimenter specifies the number of bacteria/environment combinations to be run, as well as the initial conditions for each. Data collection and storage is automated, with a computer displaying live current readings and graphing historical current levels. The experimenter can change the conditions of any fuel cell throughout the course of the experiment without affecting other fuel cells. The fuel cells themselves are stand-alone, capable of being treated as individual circuit components.<br />
<br />
===Specifications===<br />
<br />
* automated<br />
Some experiments can last several days. Measurements must be automated to allow for overnight observation. <br />
<br />
* anaerobic/aerobic<br />
S. oneidensis MR-1 only oxidizes substrates in anaerobic environments. The chambers housing the bacteria must be oxygen free and airtight. <br />
<br />
* sterile<br />
Fuel cells must be capable of being sterilized to prevent contamination<br />
<br />
* reproducible<br />
Individual fuel cells must be similar enough to produce consistent results.<br />
<br />
* accessible<br />
Experimenters must have access to the bacterial environment.<br />
<br />
==Approach==<br />
<br />
===Decomposition into components===<br />
<br />
Construction of the system was broken down into three distinct parts. The most important components were the fuel cells themselves. Once these devices were built, a measurement system was constructed to sample current readings from each simultaneously. Software was developed to orchestrate these readings, recording and displaying them in real time.<br />
<br />
===Component descriptions and approaches===<br />
<br />
====Fuel Cells====<br />
<br />
Fuel cells were constructed to provide an environment in which bacteria could live and produce electric current. After researching various microbial fuel cell designs, we concluded that a two chamber system would best suit our goals. <br />
<br />
The simple picture of our design was a hollow tube, sealed at both ends with endplates, and cut in half horizontally to allow for insertion of a semi-permeable membrane. The two halves would then be clamped back together to close the system. Ports needed to be drilled into the walls of the tube to allow for insertion of the anode and cathode, as well as to initially fill the sealed chambers with media and inject bacteria and food. Previous research suggested bubbling gasses into the chambers could improve performance. In the anode chamber, nitrogen was typically bubbled to purge the chamber of oxygen, keeping the bacteria in a controlled anaerobic environment. In the cathode chamber, air was typically bubbled to provide a continuous source of oxygen, ensuring an efficient reduction reaction. We decided to include ports for gas inlet and outlet as well.<br />
<br />
Materials for the chamber were then selected. The central tube and endplates were made from polycarbonate. This is a clear, hard plastic that is easily bonded to itself with silicone glue. It has the added benefit of allowing visual monitoring of the chambers, it is easily milled, and it can be autoclaved. Silicone was used in glue form to bond the tubes to their endplates, and it was used in solidified from to construct gaskets which sealed the two chambers together when clamped. The clamping mechanism involved drilling holes through the overlap of the endplates, inserting iron threaded rods down the length of the chambers, and fastening the ends of the rods with wing nuts.<br />
<br />
The anode material was chosen to provide maximal surface area and attachment sites for bacteria. Carbon felt provided such a surface.<br />
<br />
The cathode material needed to provide an efficient surface for the reduction reactions to take place. Platinum on carbon cloth acted as a catalyst for this reaction.<br />
<br />
Titanium wire was used to connect the cathode and anode surfaces to an external resistor as it cannot be oxidized, which would interfere with the reactions.<br />
<br />
====Measurement====<br />
<br />
We required that our measurement system be automated and capable of monitoring multiple fuel cells at once. Our original goal was to measure and compare output current levels between separate fuel cell environments. We choose to use current as the measured variable since previous studies in s. Onedisis and mutants gave us reasonable expectation values of the current magnitudes we would observe. As these reported current levels were on the order of microamps for similarly sized systems, we decided to purchase a high throughput commercial digital multimeter (DMM) to achieve the desired resolution. Finding a multiplexed system with large quantities of current channels proved difficult, and we adapted our schematic to measure voltage and resistance and then calculate current.<br />
<br />
Our final product choice was the Keithley 2700 with a Keithley 7700 multiplexer accessory. This configuration provided 100 nA current resolution while allowing up to 20 devices to be sampled from at once. <br />
<br />
We choose to use 470 Ohms as our resistance connecting the anode and cathode of each fuel cell. This choice was based on several considerations. We measured the resistance path from the surface of an anode to the surface of a cathode and found that it ranged from 5-10 Ohms. Our resistance had to be large enough that voltage drops across this internal resistance did not significantly affect the measured voltage drop across our external resistor. We also measured the resistance of the path from anode to cathode through the fuel cells media and membrane. This resistance was highly variable but typically ranged from 15K to 25K. Our external resistance had to be low enough that the vast majority of freed electrons traveled externally rather than through this variable media/membrane path.<br />
<br />
====Software====<br />
<br />
The task of our software program was to coordinate measurements through the DMM, record data, and display live data in graphical format. Some members of our team had prior experience with LabVIEW, which we used to develop these routines. <br />
<br />
Working with the Keithley DMM, our program first samples resistances across each fuel cell's external resistor. It then switches the DMM measurement channel to voltage and enters a measurement loop. In each loop iteration, voltage samples are taken from all connected fuel cells and are divided by their previously measured resistances. This computed current is displayed in both graphical and tabular format, as well as stored in a .xls file. Current values are compared against a user specified threshold value, and a display indicates which chambers are over threshold. To avoid spikes, this function only returns true if a chamber's current level has been above threshold for the previous ten readings. An iteration ends by waiting for the next multiple of a user specified time. This ensures synchronous measurement intervals.<br />
<br />
* In LabVIEW, a block diagram is used to visually write c code. Our block diagram is shown here.<br />
[[Image:Labview.jpg|600px]]<br />
<br />
<br />
* In Labview, an instrument panel is used to allow users to specify inputs and to display the status of program variables. Our instrument panel is shown here.<br />
[[Image:Labview2.jpg|600px]]<br />
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<div></div>Joyyhttp://2008.igem.org/Team:Harvard/HardwareTeam:Harvard/Hardware2008-10-30T04:10:40Z<p>Joyy: /* Software */</p>
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=Fuel Cell Development=<br />
==Motivation==<br />
<br />
The broad goal of our project was to engineer ''S. Oneidensis'' to produce a detectable change in electric current in response to some environmental stimulus. In order to observe such a reaction, our first task was to design an environment capable of housing bacteria and measuring current production.<br />
<br />
==Solution - Microbial Fuel Cells==<br />
<br />
===Background===<br />
<br />
To measure the electric current production in Shewanella oneidensis MR-1, we constructed microbial fuel cells (MFCs) to harvest the electricity produced. Traditional fuel cells convert chemical energy to electrical energy. MFCs are unique in that the fuel source used is microbially degradable organic matter (Lovley 2006). Such organic matter is a form of renewable energy and can come from materials in wastewater, sediments, or agricultural wastes (Cho et al. 2007). Compared to hydrogen- and methanol-driven fuel cells, MFCs are particular attractive due to their ease of operation and wide range of fuel sources available. They do not require expensive catalysts, high operating temperatures, nor explosive or toxic fuels (Lovley 2006). Indeed, we found the construction and operation of our microbial fuel cells to be relatively straightforward. <br />
<br />
MFCs operate using principles similar to normal fuel cells. MFCs require an anode and a cathode separated by a semi-permeable membrane (permeable to protons) (Lovley 2006). The anode compartment is generally anaerobic and is the site of oxidation of organic matter (Lovley 2006); the cathode transfers electrons to the electron acceptor (Lovley 2006). MFCs take advantage of the natural metabolism of microorganisms that break down organic matter by intercepting the electrons that would be donated to naturally occurring electron acceptors. Instead, these electrons are transferred from the anode to the cathode, usually along metal wires to which current readings can be taken. <br />
<br />
A figure of a microbial fuel cell is shown below (NB: this is not what we used in our experiments). <br />
<br />
[[ Image: Snapshot1.png | 350px ]]<br />
<br />
On the anode side, glucose is being metabolized by the microorganism to yield electrons which are transferred to the cathode side where oxygen is reduced to water (Lovley 2006).<br />
<br />
===Context===<br />
<br />
Shewanella oneidensis MR-1 is a suitable microorganism for a MFC. It is known to break down lactate (Bretschger et al. 2007), and in anaerobic conditions, S. oneidensis MR-1 produces nanowires that shuttle electrons to electron acceptors as its aerobic electron acceptor, oxygen, is unavailable (Gorby et al. 2006). Taking advantage of this, we designed our microbial fuel cell with an anaerobic anode where completion of the lactate reduction pathway must be completed by transferring electrons to the cathode side at which reduction of oxygen to water occurs. The anode chamber, besides housing our bacteria, was also the site of chemical injections. We provided S. oneidensis MR-1 with lactate as its sole carbon source and sought to induce the mtrB gene, thus “turning on” current production, with chemicals such as lactose and tetracycline. In addition, our heat tests were directed towards the anode chamber as well. Thus, the detection of electricity production of S. oneidensis MR-1 using MFCs serves as an innovative way to detect gene expression, combining both traditional molecular biology with renewable energy engineering. <br />
<br />
<br />
[[Image:Chambers.jpg|600px]]<br />
<br />
==Design Goal==<br />
<br />
===Functional description===<br />
<br />
The final product is a complete system capable of introducing separate strains of bacteria to multiple different environments while simultaneously measuring and recording current readings from each. The experimenter specifies the number of bacteria/environment combinations to be run, as well as the initial conditions for each. Data collection and storage is automated, with a computer displaying live current readings and graphing historical current levels. The experimenter can change the conditions of any fuel cell throughout the course of the experiment without affecting other fuel cells. The fuel cells themselves are stand-alone, capable of being treated as individual circuit components.<br />
<br />
===Specifications===<br />
<br />
* automated<br />
Some experiments can last several days. Measurements must be automated to allow for overnight observation. <br />
<br />
* anaerobic/aerobic<br />
S. oneidensis MR-1 only oxidizes substrates in anaerobic environments. The chambers housing the bacteria must be oxygen free and airtight. <br />
<br />
* sterile<br />
Fuel cells must be capable of being sterilized to prevent contamination<br />
<br />
* reproducible<br />
Individual fuel cells must be similar enough to produce consistent results.<br />
<br />
* accessible<br />
Experimenters must have access to the bacterial environment.<br />
<br />
==Approach==<br />
<br />
===Decomposition into components===<br />
<br />
Construction of the system was broken down into three distinct parts. The most important components were the fuel cells themselves. Once these devices were built, a measurement system was constructed to sample current readings from each simultaneously. Software was developed to orchestrate these readings, recording and displaying them in real time.<br />
<br />
===Component descriptions and approaches===<br />
<br />
====Fuel Cells====<br />
<br />
Fuel cells were constructed to provide an environment in which bacteria could live and produce electric current. After researching various microbial fuel cell designs, we concluded that a two chamber system would best suit our goals. <br />
<br />
The simple picture of our design was a hollow tube, sealed at both ends with endplates, and cut in half horizontally to allow for insertion of a semi-permeable membrane. The two halves would then be clamped back together to close the system. Ports needed to be drilled into the walls of the tube to allow for insertion of the anode and cathode, as well as to initially fill the sealed chambers with media and inject bacteria and food. Previous research suggested bubbling gasses into the chambers could improve performance. In the anode chamber, nitrogen was typically bubbled to purge the chamber of oxygen, keeping the bacteria in a controlled anaerobic environment. In the cathode chamber, air was typically bubbled to provide a continuous source of oxygen, ensuring an efficient reduction reaction. We decided to include ports for gas inlet and outlet as well.<br />
<br />
Materials for the chamber were then selected. The central tube and endplates were made from polycarbonate. This is a clear, hard plastic that is easily bonded to itself with silicone glue. It has the added benefit of allowing visual monitoring of the chambers, it is easily milled, and it can be autoclaved. Silicone was used in glue form to bond the tubes to their endplates, and it was used in solidified from to construct gaskets which sealed the two chambers together when clamped. The clamping mechanism involved drilling holes through the overlap of the endplates, inserting iron threaded rods down the length of the chambers, and fastening the ends of the rods with wing nuts.<br />
<br />
The anode material was chosen to provide maximal surface area and attachment sites for bacteria. Carbon felt provided such a surface.<br />
<br />
The cathode material needed to provide an efficient surface for the reduction reactions to take place. Platinum on carbon cloth acted as a catalyst for this reaction.<br />
<br />
Titanium wire was used to connect the cathode and anode surfaces to an external resistor as it cannot be oxidized, which would interfere with the reactions.<br />
<br />
====Measurement====<br />
<br />
We required that our measurement system be automated and capable of monitoring multiple fuel cells at once. Our original goal was to measure and compare output current levels between separate fuel cell environments. We choose to use current as the measured variable since previous studies in s. Onedisis and mutants gave us reasonable expectation values of the current magnitudes we would observe. As these reported current levels were on the order of microamps for similarly sized systems, we decided to purchase a high throughput commercial digital multimeter (DMM) to achieve the desired resolution. Finding a multiplexed system with large quantities of current channels proved difficult, and we adapted our schematic to measure voltage and resistance and then calculate current.<br />
<br />
Our final product choice was the Keithley 2700 with a Keithley 7700 multiplexer accessory. This configuration provided 100 nA current resolution while allowing up to 20 devices to be sampled from at once. <br />
<br />
We choose to use 470 Ohms as our resistance connecting the anode and cathode of each fuel cell. This choice was based on several considerations. We measured the resistance path from the surface of an anode to the surface of a cathode and found that it ranged from 5-10 Ohms. Our resistance had to be large enough that voltage drops across this internal resistance did not significantly affect the measured voltage drop across our external resistor. We also measured the resistance of the path from anode to cathode through the fuel cells media and membrane. This resistance was highly variable but typically ranged from 15K to 25K. Our external resistance had to be low enough that the vast majority of freed electrons traveled externally rather than through this variable media/membrane path.<br />
<br />
====Software====<br />
<br />
The task of our software program was to coordinate measurements through the DMM, record data, and display live data in graphical format. Some members of our team had prior experience with LabVIEW, which we used to develop these routines. <br />
<br />
Working with the Keithley DMM, our program first samples resistances across each fuel cell's external resistor. It then switches the DMM measurement channel to voltage and enters a measurement loop. In each loop iteration, voltage samples are taken from all connected fuel cells and are divided by their previously measured resistances. This computed current is displayed in both graphical and tabular format, as well as stored in a .xls file. Current values are compared against a user specified threshold value, and a display indicates which chambers are over threshold. To avoid spikes, this function only returns true if a chamber's current level has been above threshold for the previous ten readings. An iteration ends by waiting for the next multiple of a user specified time. This ensures synchronous measurement intervals.<br />
<br />
* In LabVIEW, a block diagram is used to visually write c code. Our block diagram is shown here.<br />
[[Image:Labview.jpg|600px]]<br />
<br />
<br />
* In Labview, an instrument panel is used to allow users to specify inputs and to display the status of program variables. Our instrument panel is shown here.<br />
[[Image:LV_vi1.png |350px| left]][[Image:LV_vi2.png | 350px]]<br />
<br />
|}<br />
<br><br><br />
<br />
<!--- end body ---><br />
|}</div>Joyyhttp://2008.igem.org/File:Labview.jpgFile:Labview.jpg2008-10-30T04:10:08Z<p>Joyy: </p>
<hr />
<div></div>Joyyhttp://2008.igem.org/Team:Harvard/HardwareTeam:Harvard/Hardware2008-10-30T04:07:54Z<p>Joyy: /* Software */</p>
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|<br />
=Fuel Cell Development=<br />
==Motivation==<br />
<br />
The broad goal of our project was to engineer ''S. Oneidensis'' to produce a detectable change in electric current in response to some environmental stimulus. In order to observe such a reaction, our first task was to design an environment capable of housing bacteria and measuring current production.<br />
<br />
==Solution - Microbial Fuel Cells==<br />
<br />
===Background===<br />
<br />
To measure the electric current production in Shewanella oneidensis MR-1, we constructed microbial fuel cells (MFCs) to harvest the electricity produced. Traditional fuel cells convert chemical energy to electrical energy. MFCs are unique in that the fuel source used is microbially degradable organic matter (Lovley 2006). Such organic matter is a form of renewable energy and can come from materials in wastewater, sediments, or agricultural wastes (Cho et al. 2007). Compared to hydrogen- and methanol-driven fuel cells, MFCs are particular attractive due to their ease of operation and wide range of fuel sources available. They do not require expensive catalysts, high operating temperatures, nor explosive or toxic fuels (Lovley 2006). Indeed, we found the construction and operation of our microbial fuel cells to be relatively straightforward. <br />
<br />
MFCs operate using principles similar to normal fuel cells. MFCs require an anode and a cathode separated by a semi-permeable membrane (permeable to protons) (Lovley 2006). The anode compartment is generally anaerobic and is the site of oxidation of organic matter (Lovley 2006); the cathode transfers electrons to the electron acceptor (Lovley 2006). MFCs take advantage of the natural metabolism of microorganisms that break down organic matter by intercepting the electrons that would be donated to naturally occurring electron acceptors. Instead, these electrons are transferred from the anode to the cathode, usually along metal wires to which current readings can be taken. <br />
<br />
A figure of a microbial fuel cell is shown below (NB: this is not what we used in our experiments). <br />
<br />
[[ Image: Snapshot1.png | 350px ]]<br />
<br />
On the anode side, glucose is being metabolized by the microorganism to yield electrons which are transferred to the cathode side where oxygen is reduced to water (Lovley 2006).<br />
<br />
===Context===<br />
<br />
Shewanella oneidensis MR-1 is a suitable microorganism for a MFC. It is known to break down lactate (Bretschger et al. 2007), and in anaerobic conditions, S. oneidensis MR-1 produces nanowires that shuttle electrons to electron acceptors as its aerobic electron acceptor, oxygen, is unavailable (Gorby et al. 2006). Taking advantage of this, we designed our microbial fuel cell with an anaerobic anode where completion of the lactate reduction pathway must be completed by transferring electrons to the cathode side at which reduction of oxygen to water occurs. The anode chamber, besides housing our bacteria, was also the site of chemical injections. We provided S. oneidensis MR-1 with lactate as its sole carbon source and sought to induce the mtrB gene, thus “turning on” current production, with chemicals such as lactose and tetracycline. In addition, our heat tests were directed towards the anode chamber as well. Thus, the detection of electricity production of S. oneidensis MR-1 using MFCs serves as an innovative way to detect gene expression, combining both traditional molecular biology with renewable energy engineering. <br />
<br />
<br />
[[Image:Chambers.jpg|600px]]<br />
<br />
==Design Goal==<br />
<br />
===Functional description===<br />
<br />
The final product is a complete system capable of introducing separate strains of bacteria to multiple different environments while simultaneously measuring and recording current readings from each. The experimenter specifies the number of bacteria/environment combinations to be run, as well as the initial conditions for each. Data collection and storage is automated, with a computer displaying live current readings and graphing historical current levels. The experimenter can change the conditions of any fuel cell throughout the course of the experiment without affecting other fuel cells. The fuel cells themselves are stand-alone, capable of being treated as individual circuit components.<br />
<br />
===Specifications===<br />
<br />
* automated<br />
Some experiments can last several days. Measurements must be automated to allow for overnight observation. <br />
<br />
* anaerobic/aerobic<br />
S. oneidensis MR-1 only oxidizes substrates in anaerobic environments. The chambers housing the bacteria must be oxygen free and airtight. <br />
<br />
* sterile<br />
Fuel cells must be capable of being sterilized to prevent contamination<br />
<br />
* reproducible<br />
Individual fuel cells must be similar enough to produce consistent results.<br />
<br />
* accessible<br />
Experimenters must have access to the bacterial environment.<br />
<br />
==Approach==<br />
<br />
===Decomposition into components===<br />
<br />
Construction of the system was broken down into three distinct parts. The most important components were the fuel cells themselves. Once these devices were built, a measurement system was constructed to sample current readings from each simultaneously. Software was developed to orchestrate these readings, recording and displaying them in real time.<br />
<br />
===Component descriptions and approaches===<br />
<br />
====Fuel Cells====<br />
<br />
Fuel cells were constructed to provide an environment in which bacteria could live and produce electric current. After researching various microbial fuel cell designs, we concluded that a two chamber system would best suit our goals. <br />
<br />
The simple picture of our design was a hollow tube, sealed at both ends with endplates, and cut in half horizontally to allow for insertion of a semi-permeable membrane. The two halves would then be clamped back together to close the system. Ports needed to be drilled into the walls of the tube to allow for insertion of the anode and cathode, as well as to initially fill the sealed chambers with media and inject bacteria and food. Previous research suggested bubbling gasses into the chambers could improve performance. In the anode chamber, nitrogen was typically bubbled to purge the chamber of oxygen, keeping the bacteria in a controlled anaerobic environment. In the cathode chamber, air was typically bubbled to provide a continuous source of oxygen, ensuring an efficient reduction reaction. We decided to include ports for gas inlet and outlet as well.<br />
<br />
Materials for the chamber were then selected. The central tube and endplates were made from polycarbonate. This is a clear, hard plastic that is easily bonded to itself with silicone glue. It has the added benefit of allowing visual monitoring of the chambers, it is easily milled, and it can be autoclaved. Silicone was used in glue form to bond the tubes to their endplates, and it was used in solidified from to construct gaskets which sealed the two chambers together when clamped. The clamping mechanism involved drilling holes through the overlap of the endplates, inserting iron threaded rods down the length of the chambers, and fastening the ends of the rods with wing nuts.<br />
<br />
The anode material was chosen to provide maximal surface area and attachment sites for bacteria. Carbon felt provided such a surface.<br />
<br />
The cathode material needed to provide an efficient surface for the reduction reactions to take place. Platinum on carbon cloth acted as a catalyst for this reaction.<br />
<br />
Titanium wire was used to connect the cathode and anode surfaces to an external resistor as it cannot be oxidized, which would interfere with the reactions.<br />
<br />
====Measurement====<br />
<br />
We required that our measurement system be automated and capable of monitoring multiple fuel cells at once. Our original goal was to measure and compare output current levels between separate fuel cell environments. We choose to use current as the measured variable since previous studies in s. Onedisis and mutants gave us reasonable expectation values of the current magnitudes we would observe. As these reported current levels were on the order of microamps for similarly sized systems, we decided to purchase a high throughput commercial digital multimeter (DMM) to achieve the desired resolution. Finding a multiplexed system with large quantities of current channels proved difficult, and we adapted our schematic to measure voltage and resistance and then calculate current.<br />
<br />
Our final product choice was the Keithley 2700 with a Keithley 7700 multiplexer accessory. This configuration provided 100 nA current resolution while allowing up to 20 devices to be sampled from at once. <br />
<br />
We choose to use 470 Ohms as our resistance connecting the anode and cathode of each fuel cell. This choice was based on several considerations. We measured the resistance path from the surface of an anode to the surface of a cathode and found that it ranged from 5-10 Ohms. Our resistance had to be large enough that voltage drops across this internal resistance did not significantly affect the measured voltage drop across our external resistor. We also measured the resistance of the path from anode to cathode through the fuel cells media and membrane. This resistance was highly variable but typically ranged from 15K to 25K. Our external resistance had to be low enough that the vast majority of freed electrons traveled externally rather than through this variable media/membrane path.<br />
<br />
====Software====<br />
<br />
The task of our software program was to coordinate measurements through the DMM, record data, and display live data in graphical format. Some members of our team had prior experience with LabVIEW, which we used to develop these routines. <br />
<br />
Working with the Keithley DMM, our program first samples resistances across each fuel cell's external resistor. It then switches the DMM measurement channel to voltage and enters a measurement loop. In each loop iteration, voltage samples are taken from all connected fuel cells and are divided by their previously measured resistances. This computed current is displayed in both graphical and tabular format, as well as stored in a .xls file. Current values are compared against a user specified threshold value, and a display indicates which chambers are over threshold. To avoid spikes, this function only returns true if a chamber's current level has been above threshold for the previous ten readings. An iteration ends by waiting for the next multiple of a user specified time. This ensures synchronous measurement intervals.<br />
<br />
* In LabVIEW, a block diagram is used to visually write c code. Our block diagram is shown here.<br />
[[Image:LV_block1.png |350px|left]][[Image:LV_block2.png | 350px|]]<br />
<br />
<br />
* In Labview, an instrument panel is used to allow users to specify inputs and to display the status of program variables. Our instrument panel is shown here.<br />
[[Image:LV_vi1.png |350px| left]][[Image:LV_vi2.png | 350px]]<br />
<br />
|}<br />
<br><br><br />
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<!--- end body ---><br />
|}</div>Joyyhttp://2008.igem.org/Team:Harvard/Hardware/MFCProcedureTeam:Harvard/Hardware/MFCProcedure2008-10-30T04:05:38Z<p>Joyy: /* Gas Tubing Assembly */</p>
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=Running an MFC Experiment=<br />
<br />
This page is intended as a comprehensive guide to completing a microbial fuel cell setup and running an experiment from start to finish. <br />
<br />
==Creating a Testing Environment==<br />
<br />
Begin 1-2 weeks prior to experiment<br />
<br />
===Constructing Fuel Cell Components===<br />
<br />
'''Materials (per fuel cell)'''<br />
<br />
* 4" Polycarbonate Square Tube, 2" Outer Diameter<br />
* 6" x 6" Polycarbonate Sheet, 1/4" Thick<br />
* 4 Steel Fully Threaded Stud, 1/4"-20 Thread, 6" Length<br />
* 8 Zinc Alloy Wing Flange Nut, 1/4"-20 Screw Size, 1" Wing Spread<br />
* 1" x 1" Nafion® membrane, 0.180mm thick<br />
* 1" x 1" Carbon felt, 0.25" thick<br />
* 1.5" x 1.5" E-TEK ELAT™ GDE (platinum on carbon)<br />
* 2' Titanium Grade 2 Wire .046" Diameter<br />
* Teflon Tape, 1/4" Width<br />
* 5" x 2.5" Silicone Sheet<br />
* Silicone Glue<br />
* Spiral Point Tap 1/4"-28<br />
* 8 Plastic Luer Lock Coupling Nylon, Female to Male Thread, 1/4"-28<br />
* 8 Luer Lock Injection Ports<br />
<br />
'''Procedure'''<br />
<br />
1) Mill Polycarbonate<br />
* Cut polycarbonate sheet into 4 equal 3" x 3" pieces<br />
* Drill four 3/8" holes through each piece, 1 per corner, indented 5mm from both sides<br />
[[Image:endplates.jpg | 600px]]<br />
* Cut polycarbonate tube into two equal 2" halves<br />
* Drill four 1/4" holes through each half in configuration shown<br />
[[Image:drilled_tube.jpg | 600px]]<br />
* Tap each hole with 1/4" -28 spiral tap<br />
<br />
2) Glue Chambers (repeat for each half)<br />
* Center tube on endplate by marking plate with 'X' from corner to corner<br />
* Squirt 2mm thick line of silicone on edge of tube (edge furthest from holes)<br />
* Press tube firmly against marked location on endplate<br />
* Quickly spread excess silicone along edge<br />
* Let stand 24h to harden<br />
[[Image:glued_half.jpg | 600px]]<br />
<br />
3) Construct Gaskets<br />
* Cut silicone sheet into two equal 2.25" x 2.25" pieces<br />
* Cut out centered inner squares in each piece, 1.75" x 1.75"<br />
* Using inner squares, cut two 'O' rings, inner diameter 1/4", outer diameter 1/2"<br />
[[Image:gaskets.jpg | 600px]]<br />
<br />
4) Construct Electrodes<br />
* Cut titanium wire into one 8" piece and one 16" piece<br />
* Using pliers, shape anode and cathode as shown<br />
'''Anode Frame'''<br><br />
[[Image:Anodeframe.jpg | 500px]]<br><br />
'''Cathode Frame'''<br><br />
[[Image:Cathodeframe.jpg | 500px]]<br />
<br />
* Spear carbon felt with tip of anode titanium wire and wedge into frame<br />
* Weave platinum carbon cloth through cathode titanium wire<br />
'''Anode'''<br><br />
[[Image:Anode_electrode.jpg | 500px]]<br><br />
'''Cathode'''<br><br />
[[Image:Cathode_electrode.jpg | 500px]]<br />
<br />
5) Seal Injection Ports<br />
* Wrap threads of all eight Luer Lock screws with 1' of teflon tape in opposite direction of screwing<br />
* Screw Luer Locks into all tapped holes in both chambers<br />
[[Image:fin_chamber.jpg | 600px]]<br />
<br />
===Setup of Measurement Device===<br />
<br />
'''Materials'''<br />
* Keithley 2700 Digital Multimeter<br />
* Keithley 7700 Multiplexer<br />
* Small Breadboard<br />
* Supply of insulted thin copper wire<br />
* 470 Ohm resistors (one/fuel cell)<br />
<br />
'''Procedure'''<br />
1) Wire Multiplexer<br />
* Open multiplexer, note channels<br />
* Cut two wire 18" wire leads per fuel cell<br />
* Strip ends, place one wire in each screw terminal, screw tight<br />
* Tape paired wires (two are attached to each channel) near non-attached ends and label<br />
* Clamp wire bundles near back of device with provided plastic latch clamps<br />
* Close Multiplexer and slide into 2700 DMM<br />
<br />
2) Create Resistor Array<br />
* Connect resistors across middle of breadboard (one per fuel cell)<br />
* Connect leads from multiplexer across resistors (one pair across each resistor)<br />
<br />
<br />
===Controlling the DMM with LabVIEW™===<br />
<br />
# Initialize Multimeter<br />
#* Attach 2700 to COM1 port of desktop computer w/ LabVIEW™<br />
#* Download our LabVIEW™ source code [[Media:MFCs.txt|MFCs.vi]]<br />
#* Open Program in LabVIEW™, adjust block diagram as necessary<br />
<br />
==Experiment Preparation==<br />
<br />
Begin 1 day prior to experiment<br />
<br />
===Assembling Chambers===<br />
<br />
'''Procedure'''<br><br />
1) Prepare Electrodes<br />
* Attach Luer Lock injection ports to all chamber screws<br />
* Poke tip of electrodes through designated ports from the inside<br />
[[Image:chamber_w_elec.jpg | 600px]]<br />
<br><br />
2) Align Chambers<br />
* Lay one chamber on a flat surface<br />
* Place silicone square ring on top edge of tube<br />
* Place polycarbonate square on silicone<br />
* Place silicone 'O' ring around central pore<br />
* Place Nafion membrane on top of 'O' ring<br />
* Sandwich membrane between second 'O' ring<br />
* Align second polycarbonate square on top of 'O' ring<br />
* Center second silicone square ring on polycarbonate square<br />
* Set second chamber on top of silicone, ensuring ports facing same direction as first chamber<br />
<br><br />
3) Clamp Chambers<br />
* Move assembly into vice or clamp<br />
* Insert rods through holes in end plates and screw on wing nuts<br />
* Tighten evenly<br />
<br />
===Solutions Prep===<br />
<br />
# Chamber media (150ml / fuel cell)<br />
#* 5.844 g/L 100mM NaCl<br />
#* 15.1185 g/L 50mM PIPES (hydrogen) <br />
#* ''7.0 pH''<br />
# Phosphate buffer (60ml / fuel cell)<br />
#* 2.918 g/L Monosodium phosphate, monohydrate<br />
#* 4.095 g/L Disodium phosphate, anhydrous <br />
#* 5.844 g/L 100mM NaCl<br />
#* ''7.0 pH''<br />
<br />
===Gas Tubing Assembly===<br />
<br />
'''Materials'''<br />
* 25' Silicone Soft Rubber Tubing, 3/32" ID, 7/32" OD, 1/16" Wall<br />
* Tank of Compressed Nitrogen<br />
* Gas Regulator<br />
* Lab Supply of Air<br />
* 4 Plastic Luer Lock Coupling Nylon, Male X Barb, for 3/32" Tube<br />
* 4 Plastic Luer Lock Coupling Nylon, Female X Barb, for 3/32" Tube<br />
* Plastic Luer Lock Coupling Nylon, T junctions, for 3/32" Tube<br />
* Syringe needles - 27 gauge<br />
* 2 Aspirator Flasks<br />
* 2 Rubber Stoppers<br />
<br />
'''Procedure'''<br />
1) Make Flow Regulators<br />
* Insert nozzle of female Luer Lock into rubber stoppers (poke hole if necessary)<br />
* Cap aspirator flasks with rubber stoppers<br />
* Attach tubing from gas sources to each glass nozzle of aspirator flask<br />
[[Image:flow_regs.jpg | 600px]]<br />
<br />
2) Make Manifolds<br />
* Attach T-junction Luer Lock pieces into manifold (1 junction/ fuel cell ; 2 manifods total)<br />
* Turn last juction such that off is facing end of manifold<br />
* Attach tubing from stopper of flow regulators to beginning of each mainfold<br />
[[Image:manifolds.jpg | 600px]]<br />
<br />
===Growing Strains===<br />
<br />
Materials<br />
<br />
* 150mL LB / strain<br />
* 1000mL airating flask / strain<br />
* Antibiotics (if desired strain must be selected for)<br />
* Plate or Glycerol Stock with desired strain<br />
<br />
Procedure<br />
<br />
# Fill flasks with LB<br />
# Add correct concentration of selection antibiotic<br />
# Pick single colony from plate, add to flask<br />
# Incubate overnight in shaker at 30C<br />
<br />
==Runtime==<br />
<br />
Begin 2 hours prior to experiment<br />
<br />
===Bacteria===<br />
<br />
Procedure<br />
<br />
# Pipet the cells out of the flask and into 250mL centrifuge containers <br />
# Make sure the containers are close in weight (within 0.5g of each other)<br />
# For a culture more than 250mL split it into two containers <br />
# Set the centrifuge temperature to 22-23C, spin speed to 5000RPM, and time to 15 min<br />
# After first spin, drain each container of the LB, making sure to leave the bacteria pellet intact<br />
# Resuspend pellet in 50mL of potassium buffer<br />
# After bacteria are fully resuspended (no pellets at all), spin down again, 22-23C, 5000RPM, 15min.<br />
# Pour potassium buffer out of container slowly.<br />
# Resuspend pellet in 50mL of potassium buffer<br />
# Spin down again, 22-23C, 5000RPM, 15min.<br />
# Pour potassium buffer out of container slowly. <br />
# Resuspend pellet in 4 mL of sodium pipes<br />
# Check OD (100microliters in 15mL or 1:150 dilution)<br />
# If OD is in linear range, calculate dilution for desired quantity of bacteria in 1mL (typically 10^8 cells /mL of chamber media)<br />
# Repeat dilution if not in linear range (using different ratio)<br />
# Inject 1mL of bacteria into each chamber (see below)<br />
<br />
===Fuel Cells===<br />
<br />
Procedure<br />
<br />
# Pipet 75mL of NaPiPES solution into each side of fuel cells<br />
# Inject 1mL of Resazurin solution into each side of chamber<br />
# Using Luer Lock nozzles, connect tubing from top ports to beaker w/ distilled water<br />
# Cut tubing to span distance from manifolds to each fuel cell (1 from nitrogen, 1 from air)<br />
# Attach syringe needles to tubing via Luer Lock nozzles<br />
# Using Luer Lock nozzles, connect tubing from manifolds<br />
# Start gas flow<br />
# Poke needles through bottom ports on fuel cells<br />
<br />
===Measurements===<br />
<br />
Procedure<br />
<br />
# Turn on computer and digital multimeter<br />
# Open LabVIEW program<br />
# Click Run arrow to take resistance readings<br />
# Connect fuel cells to resistor array via alligator clips<br />
# Click "Begin Current Readings" Icon on instrument display<br />
<br />
===Injections/Variables===<br />
<br />
Procedure<br />
<br />
# Once current readings reach equilibrium, inject bacteria into fuels cells using syringes<br />
# Allow bacteria to consume any carbon sources left in their media (approximately 12 hours)<br />
# Once current levels reach stable baseline, inject 1ml lactate solution<br />
# Inject additional variables as desired<br />
<br />
==Clean Up==<br />
<br />
Procedure<br />
<br />
# Drain chambers and soak in 70% ethanol<br />
# Remove carbon felt from anodes and discard (save titanium)<br />
# Scrub all parts in ethanol and distilled water successively<br />
# Use pipe cleaners on ports and tubes<br />
<br />
<br />
<br />
|}<br />
<br><br><br />
<br />
<!--- end body ---><br />
|}</div>Joyyhttp://2008.igem.org/Team:Harvard/Hardware/MFCProcedureTeam:Harvard/Hardware/MFCProcedure2008-10-30T04:04:59Z<p>Joyy: /* Solutions Prep */</p>
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<!--- body here---><br />
{|align="justify" style="background-color:#FFFFFF;text-indent: 15pt;text-align:justify" cellpadding="50" width="90%"<br />
|<br />
=Running an MFC Experiment=<br />
<br />
This page is intended as a comprehensive guide to completing a microbial fuel cell setup and running an experiment from start to finish. <br />
<br />
==Creating a Testing Environment==<br />
<br />
Begin 1-2 weeks prior to experiment<br />
<br />
===Constructing Fuel Cell Components===<br />
<br />
'''Materials (per fuel cell)'''<br />
<br />
* 4" Polycarbonate Square Tube, 2" Outer Diameter<br />
* 6" x 6" Polycarbonate Sheet, 1/4" Thick<br />
* 4 Steel Fully Threaded Stud, 1/4"-20 Thread, 6" Length<br />
* 8 Zinc Alloy Wing Flange Nut, 1/4"-20 Screw Size, 1" Wing Spread<br />
* 1" x 1" Nafion® membrane, 0.180mm thick<br />
* 1" x 1" Carbon felt, 0.25" thick<br />
* 1.5" x 1.5" E-TEK ELAT™ GDE (platinum on carbon)<br />
* 2' Titanium Grade 2 Wire .046" Diameter<br />
* Teflon Tape, 1/4" Width<br />
* 5" x 2.5" Silicone Sheet<br />
* Silicone Glue<br />
* Spiral Point Tap 1/4"-28<br />
* 8 Plastic Luer Lock Coupling Nylon, Female to Male Thread, 1/4"-28<br />
* 8 Luer Lock Injection Ports<br />
<br />
'''Procedure'''<br />
<br />
1) Mill Polycarbonate<br />
* Cut polycarbonate sheet into 4 equal 3" x 3" pieces<br />
* Drill four 3/8" holes through each piece, 1 per corner, indented 5mm from both sides<br />
[[Image:endplates.jpg | 600px]]<br />
* Cut polycarbonate tube into two equal 2" halves<br />
* Drill four 1/4" holes through each half in configuration shown<br />
[[Image:drilled_tube.jpg | 600px]]<br />
* Tap each hole with 1/4" -28 spiral tap<br />
<br />
2) Glue Chambers (repeat for each half)<br />
* Center tube on endplate by marking plate with 'X' from corner to corner<br />
* Squirt 2mm thick line of silicone on edge of tube (edge furthest from holes)<br />
* Press tube firmly against marked location on endplate<br />
* Quickly spread excess silicone along edge<br />
* Let stand 24h to harden<br />
[[Image:glued_half.jpg | 600px]]<br />
<br />
3) Construct Gaskets<br />
* Cut silicone sheet into two equal 2.25" x 2.25" pieces<br />
* Cut out centered inner squares in each piece, 1.75" x 1.75"<br />
* Using inner squares, cut two 'O' rings, inner diameter 1/4", outer diameter 1/2"<br />
[[Image:gaskets.jpg | 600px]]<br />
<br />
4) Construct Electrodes<br />
* Cut titanium wire into one 8" piece and one 16" piece<br />
* Using pliers, shape anode and cathode as shown<br />
'''Anode Frame'''<br><br />
[[Image:Anodeframe.jpg | 500px]]<br><br />
'''Cathode Frame'''<br><br />
[[Image:Cathodeframe.jpg | 500px]]<br />
<br />
* Spear carbon felt with tip of anode titanium wire and wedge into frame<br />
* Weave platinum carbon cloth through cathode titanium wire<br />
'''Anode'''<br><br />
[[Image:Anode_electrode.jpg | 500px]]<br><br />
'''Cathode'''<br><br />
[[Image:Cathode_electrode.jpg | 500px]]<br />
<br />
5) Seal Injection Ports<br />
* Wrap threads of all eight Luer Lock screws with 1' of teflon tape in opposite direction of screwing<br />
* Screw Luer Locks into all tapped holes in both chambers<br />
[[Image:fin_chamber.jpg | 600px]]<br />
<br />
===Setup of Measurement Device===<br />
<br />
'''Materials'''<br />
* Keithley 2700 Digital Multimeter<br />
* Keithley 7700 Multiplexer<br />
* Small Breadboard<br />
* Supply of insulted thin copper wire<br />
* 470 Ohm resistors (one/fuel cell)<br />
<br />
'''Procedure'''<br />
1) Wire Multiplexer<br />
* Open multiplexer, note channels<br />
* Cut two wire 18" wire leads per fuel cell<br />
* Strip ends, place one wire in each screw terminal, screw tight<br />
* Tape paired wires (two are attached to each channel) near non-attached ends and label<br />
* Clamp wire bundles near back of device with provided plastic latch clamps<br />
* Close Multiplexer and slide into 2700 DMM<br />
<br />
2) Create Resistor Array<br />
* Connect resistors across middle of breadboard (one per fuel cell)<br />
* Connect leads from multiplexer across resistors (one pair across each resistor)<br />
<br />
<br />
===Controlling the DMM with LabVIEW™===<br />
<br />
# Initialize Multimeter<br />
#* Attach 2700 to COM1 port of desktop computer w/ LabVIEW™<br />
#* Download our LabVIEW™ source code [[Media:MFCs.txt|MFCs.vi]]<br />
#* Open Program in LabVIEW™, adjust block diagram as necessary<br />
<br />
==Experiment Preparation==<br />
<br />
Begin 1 day prior to experiment<br />
<br />
===Assembling Chambers===<br />
<br />
'''Procedure'''<br><br />
1) Prepare Electrodes<br />
* Attach Luer Lock injection ports to all chamber screws<br />
* Poke tip of electrodes through designated ports from the inside<br />
[[Image:chamber_w_elec.jpg | 600px]]<br />
<br><br />
2) Align Chambers<br />
* Lay one chamber on a flat surface<br />
* Place silicone square ring on top edge of tube<br />
* Place polycarbonate square on silicone<br />
* Place silicone 'O' ring around central pore<br />
* Place Nafion membrane on top of 'O' ring<br />
* Sandwich membrane between second 'O' ring<br />
* Align second polycarbonate square on top of 'O' ring<br />
* Center second silicone square ring on polycarbonate square<br />
* Set second chamber on top of silicone, ensuring ports facing same direction as first chamber<br />
<br><br />
3) Clamp Chambers<br />
* Move assembly into vice or clamp<br />
* Insert rods through holes in end plates and screw on wing nuts<br />
* Tighten evenly<br />
<br />
===Solutions Prep===<br />
<br />
# Chamber media (150ml / fuel cell)<br />
#* 5.844 g/L 100mM NaCl<br />
#* 15.1185 g/L 50mM PIPES (hydrogen) <br />
#* ''7.0 pH''<br />
# Phosphate buffer (60ml / fuel cell)<br />
#* 2.918 g/L Monosodium phosphate, monohydrate<br />
#* 4.095 g/L Disodium phosphate, anhydrous <br />
#* 5.844 g/L 100mM NaCl<br />
#* ''7.0 pH''<br />
<br />
===Gas Tubing Assembly===<br />
<br />
Materials<br />
* 25' Silicone Soft Rubber Tubing, 3/32" ID, 7/32" OD, 1/16" Wall<br />
* Tank of Compressed Nitrogen<br />
* Gas Regulator<br />
* Lab Supply of Air<br />
* 4 Plastic Luer Lock Coupling Nylon, Male X Barb, for 3/32" Tube<br />
* 4 Plastic Luer Lock Coupling Nylon, Female X Barb, for 3/32" Tube<br />
* Plastic Luer Lock Coupling Nylon, T junctions, for 3/32" Tube<br />
* Syringe needles - 27 gauge<br />
* 2 Aspirator Flasks<br />
* 2 Rubber Stoppers<br />
<br />
Procedure<br />
# Make Flow Regulators<br />
#* Insert nozzle of female Luer Lock into rubber stoppers (poke hole if necessary)<br />
#* Cap aspirator flasks with rubber stoppers<br />
#* Attach tubing from gas sources to each glass nozzle of aspirator flask<br />
[[Image:flow_regs.jpg | 600px]]<br />
<br />
# Make Manifolds<br />
#* Attach T-junction Luer Lock pieces into manifold (1 junction/ fuel cell ; 2 manifods total)<br />
#* Turn last juction such that off is facing end of manifold<br />
#* Attach tubing from stopper of flow regulators to beginning of each mainfold<br />
[[Image:manifolds.jpg | 600px]]<br />
<br />
===Growing Strains===<br />
<br />
Materials<br />
<br />
* 150mL LB / strain<br />
* 1000mL airating flask / strain<br />
* Antibiotics (if desired strain must be selected for)<br />
* Plate or Glycerol Stock with desired strain<br />
<br />
Procedure<br />
<br />
# Fill flasks with LB<br />
# Add correct concentration of selection antibiotic<br />
# Pick single colony from plate, add to flask<br />
# Incubate overnight in shaker at 30C<br />
<br />
==Runtime==<br />
<br />
Begin 2 hours prior to experiment<br />
<br />
===Bacteria===<br />
<br />
Procedure<br />
<br />
# Pipet the cells out of the flask and into 250mL centrifuge containers <br />
# Make sure the containers are close in weight (within 0.5g of each other)<br />
# For a culture more than 250mL split it into two containers <br />
# Set the centrifuge temperature to 22-23C, spin speed to 5000RPM, and time to 15 min<br />
# After first spin, drain each container of the LB, making sure to leave the bacteria pellet intact<br />
# Resuspend pellet in 50mL of potassium buffer<br />
# After bacteria are fully resuspended (no pellets at all), spin down again, 22-23C, 5000RPM, 15min.<br />
# Pour potassium buffer out of container slowly.<br />
# Resuspend pellet in 50mL of potassium buffer<br />
# Spin down again, 22-23C, 5000RPM, 15min.<br />
# Pour potassium buffer out of container slowly. <br />
# Resuspend pellet in 4 mL of sodium pipes<br />
# Check OD (100microliters in 15mL or 1:150 dilution)<br />
# If OD is in linear range, calculate dilution for desired quantity of bacteria in 1mL (typically 10^8 cells /mL of chamber media)<br />
# Repeat dilution if not in linear range (using different ratio)<br />
# Inject 1mL of bacteria into each chamber (see below)<br />
<br />
===Fuel Cells===<br />
<br />
Procedure<br />
<br />
# Pipet 75mL of NaPiPES solution into each side of fuel cells<br />
# Inject 1mL of Resazurin solution into each side of chamber<br />
# Using Luer Lock nozzles, connect tubing from top ports to beaker w/ distilled water<br />
# Cut tubing to span distance from manifolds to each fuel cell (1 from nitrogen, 1 from air)<br />
# Attach syringe needles to tubing via Luer Lock nozzles<br />
# Using Luer Lock nozzles, connect tubing from manifolds<br />
# Start gas flow<br />
# Poke needles through bottom ports on fuel cells<br />
<br />
===Measurements===<br />
<br />
Procedure<br />
<br />
# Turn on computer and digital multimeter<br />
# Open LabVIEW program<br />
# Click Run arrow to take resistance readings<br />
# Connect fuel cells to resistor array via alligator clips<br />
# Click "Begin Current Readings" Icon on instrument display<br />
<br />
===Injections/Variables===<br />
<br />
Procedure<br />
<br />
# Once current readings reach equilibrium, inject bacteria into fuels cells using syringes<br />
# Allow bacteria to consume any carbon sources left in their media (approximately 12 hours)<br />
# Once current levels reach stable baseline, inject 1ml lactate solution<br />
# Inject additional variables as desired<br />
<br />
==Clean Up==<br />
<br />
Procedure<br />
<br />
# Drain chambers and soak in 70% ethanol<br />
# Remove carbon felt from anodes and discard (save titanium)<br />
# Scrub all parts in ethanol and distilled water successively<br />
# Use pipe cleaners on ports and tubes<br />
<br />
<br />
<br />
|}<br />
<br><br><br />
<br />
<!--- end body ---><br />
|}</div>Joyyhttp://2008.igem.org/Team:Harvard/Hardware/MFCProcedureTeam:Harvard/Hardware/MFCProcedure2008-10-30T04:04:46Z<p>Joyy: /* Assembling Chambers */</p>
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<!--- body here---><br />
{|align="justify" style="background-color:#FFFFFF;text-indent: 15pt;text-align:justify" cellpadding="50" width="90%"<br />
|<br />
=Running an MFC Experiment=<br />
<br />
This page is intended as a comprehensive guide to completing a microbial fuel cell setup and running an experiment from start to finish. <br />
<br />
==Creating a Testing Environment==<br />
<br />
Begin 1-2 weeks prior to experiment<br />
<br />
===Constructing Fuel Cell Components===<br />
<br />
'''Materials (per fuel cell)'''<br />
<br />
* 4" Polycarbonate Square Tube, 2" Outer Diameter<br />
* 6" x 6" Polycarbonate Sheet, 1/4" Thick<br />
* 4 Steel Fully Threaded Stud, 1/4"-20 Thread, 6" Length<br />
* 8 Zinc Alloy Wing Flange Nut, 1/4"-20 Screw Size, 1" Wing Spread<br />
* 1" x 1" Nafion® membrane, 0.180mm thick<br />
* 1" x 1" Carbon felt, 0.25" thick<br />
* 1.5" x 1.5" E-TEK ELAT™ GDE (platinum on carbon)<br />
* 2' Titanium Grade 2 Wire .046" Diameter<br />
* Teflon Tape, 1/4" Width<br />
* 5" x 2.5" Silicone Sheet<br />
* Silicone Glue<br />
* Spiral Point Tap 1/4"-28<br />
* 8 Plastic Luer Lock Coupling Nylon, Female to Male Thread, 1/4"-28<br />
* 8 Luer Lock Injection Ports<br />
<br />
'''Procedure'''<br />
<br />
1) Mill Polycarbonate<br />
* Cut polycarbonate sheet into 4 equal 3" x 3" pieces<br />
* Drill four 3/8" holes through each piece, 1 per corner, indented 5mm from both sides<br />
[[Image:endplates.jpg | 600px]]<br />
* Cut polycarbonate tube into two equal 2" halves<br />
* Drill four 1/4" holes through each half in configuration shown<br />
[[Image:drilled_tube.jpg | 600px]]<br />
* Tap each hole with 1/4" -28 spiral tap<br />
<br />
2) Glue Chambers (repeat for each half)<br />
* Center tube on endplate by marking plate with 'X' from corner to corner<br />
* Squirt 2mm thick line of silicone on edge of tube (edge furthest from holes)<br />
* Press tube firmly against marked location on endplate<br />
* Quickly spread excess silicone along edge<br />
* Let stand 24h to harden<br />
[[Image:glued_half.jpg | 600px]]<br />
<br />
3) Construct Gaskets<br />
* Cut silicone sheet into two equal 2.25" x 2.25" pieces<br />
* Cut out centered inner squares in each piece, 1.75" x 1.75"<br />
* Using inner squares, cut two 'O' rings, inner diameter 1/4", outer diameter 1/2"<br />
[[Image:gaskets.jpg | 600px]]<br />
<br />
4) Construct Electrodes<br />
* Cut titanium wire into one 8" piece and one 16" piece<br />
* Using pliers, shape anode and cathode as shown<br />
'''Anode Frame'''<br><br />
[[Image:Anodeframe.jpg | 500px]]<br><br />
'''Cathode Frame'''<br><br />
[[Image:Cathodeframe.jpg | 500px]]<br />
<br />
* Spear carbon felt with tip of anode titanium wire and wedge into frame<br />
* Weave platinum carbon cloth through cathode titanium wire<br />
'''Anode'''<br><br />
[[Image:Anode_electrode.jpg | 500px]]<br><br />
'''Cathode'''<br><br />
[[Image:Cathode_electrode.jpg | 500px]]<br />
<br />
5) Seal Injection Ports<br />
* Wrap threads of all eight Luer Lock screws with 1' of teflon tape in opposite direction of screwing<br />
* Screw Luer Locks into all tapped holes in both chambers<br />
[[Image:fin_chamber.jpg | 600px]]<br />
<br />
===Setup of Measurement Device===<br />
<br />
'''Materials'''<br />
* Keithley 2700 Digital Multimeter<br />
* Keithley 7700 Multiplexer<br />
* Small Breadboard<br />
* Supply of insulted thin copper wire<br />
* 470 Ohm resistors (one/fuel cell)<br />
<br />
'''Procedure'''<br />
1) Wire Multiplexer<br />
* Open multiplexer, note channels<br />
* Cut two wire 18" wire leads per fuel cell<br />
* Strip ends, place one wire in each screw terminal, screw tight<br />
* Tape paired wires (two are attached to each channel) near non-attached ends and label<br />
* Clamp wire bundles near back of device with provided plastic latch clamps<br />
* Close Multiplexer and slide into 2700 DMM<br />
<br />
2) Create Resistor Array<br />
* Connect resistors across middle of breadboard (one per fuel cell)<br />
* Connect leads from multiplexer across resistors (one pair across each resistor)<br />
<br />
<br />
===Controlling the DMM with LabVIEW™===<br />
<br />
# Initialize Multimeter<br />
#* Attach 2700 to COM1 port of desktop computer w/ LabVIEW™<br />
#* Download our LabVIEW™ source code [[Media:MFCs.txt|MFCs.vi]]<br />
#* Open Program in LabVIEW™, adjust block diagram as necessary<br />
<br />
==Experiment Preparation==<br />
<br />
Begin 1 day prior to experiment<br />
<br />
===Assembling Chambers===<br />
<br />
'''Procedure'''<br><br />
1) Prepare Electrodes<br />
* Attach Luer Lock injection ports to all chamber screws<br />
* Poke tip of electrodes through designated ports from the inside<br />
[[Image:chamber_w_elec.jpg | 600px]]<br />
<br><br />
2) Align Chambers<br />
* Lay one chamber on a flat surface<br />
* Place silicone square ring on top edge of tube<br />
* Place polycarbonate square on silicone<br />
* Place silicone 'O' ring around central pore<br />
* Place Nafion membrane on top of 'O' ring<br />
* Sandwich membrane between second 'O' ring<br />
* Align second polycarbonate square on top of 'O' ring<br />
* Center second silicone square ring on polycarbonate square<br />
* Set second chamber on top of silicone, ensuring ports facing same direction as first chamber<br />
<br><br />
3) Clamp Chambers<br />
* Move assembly into vice or clamp<br />
* Insert rods through holes in end plates and screw on wing nuts<br />
* Tighten evenly<br />
<br />
===Solutions Prep===<br />
<br />
# Chamber media (150ml / fuel cell)<br />
#* 5.844 g/L 100mM NaCl<br />
#* 15.1185 g/L 50mM PIPES (hydrogen) <br />
#* ''7.0 pH''<br />
<br />
# Phosphate buffer (60ml / fuel cell)<br />
#* 2.918 g/L Monosodium phosphate, monohydrate<br />
#* 4.095 g/L Disodium phosphate, anhydrous <br />
#* 5.844 g/L 100mM NaCl<br />
#* ''7.0 pH''<br />
<br />
===Gas Tubing Assembly===<br />
<br />
Materials<br />
* 25' Silicone Soft Rubber Tubing, 3/32" ID, 7/32" OD, 1/16" Wall<br />
* Tank of Compressed Nitrogen<br />
* Gas Regulator<br />
* Lab Supply of Air<br />
* 4 Plastic Luer Lock Coupling Nylon, Male X Barb, for 3/32" Tube<br />
* 4 Plastic Luer Lock Coupling Nylon, Female X Barb, for 3/32" Tube<br />
* Plastic Luer Lock Coupling Nylon, T junctions, for 3/32" Tube<br />
* Syringe needles - 27 gauge<br />
* 2 Aspirator Flasks<br />
* 2 Rubber Stoppers<br />
<br />
Procedure<br />
# Make Flow Regulators<br />
#* Insert nozzle of female Luer Lock into rubber stoppers (poke hole if necessary)<br />
#* Cap aspirator flasks with rubber stoppers<br />
#* Attach tubing from gas sources to each glass nozzle of aspirator flask<br />
[[Image:flow_regs.jpg | 600px]]<br />
<br />
# Make Manifolds<br />
#* Attach T-junction Luer Lock pieces into manifold (1 junction/ fuel cell ; 2 manifods total)<br />
#* Turn last juction such that off is facing end of manifold<br />
#* Attach tubing from stopper of flow regulators to beginning of each mainfold<br />
[[Image:manifolds.jpg | 600px]]<br />
<br />
===Growing Strains===<br />
<br />
Materials<br />
<br />
* 150mL LB / strain<br />
* 1000mL airating flask / strain<br />
* Antibiotics (if desired strain must be selected for)<br />
* Plate or Glycerol Stock with desired strain<br />
<br />
Procedure<br />
<br />
# Fill flasks with LB<br />
# Add correct concentration of selection antibiotic<br />
# Pick single colony from plate, add to flask<br />
# Incubate overnight in shaker at 30C<br />
<br />
==Runtime==<br />
<br />
Begin 2 hours prior to experiment<br />
<br />
===Bacteria===<br />
<br />
Procedure<br />
<br />
# Pipet the cells out of the flask and into 250mL centrifuge containers <br />
# Make sure the containers are close in weight (within 0.5g of each other)<br />
# For a culture more than 250mL split it into two containers <br />
# Set the centrifuge temperature to 22-23C, spin speed to 5000RPM, and time to 15 min<br />
# After first spin, drain each container of the LB, making sure to leave the bacteria pellet intact<br />
# Resuspend pellet in 50mL of potassium buffer<br />
# After bacteria are fully resuspended (no pellets at all), spin down again, 22-23C, 5000RPM, 15min.<br />
# Pour potassium buffer out of container slowly.<br />
# Resuspend pellet in 50mL of potassium buffer<br />
# Spin down again, 22-23C, 5000RPM, 15min.<br />
# Pour potassium buffer out of container slowly. <br />
# Resuspend pellet in 4 mL of sodium pipes<br />
# Check OD (100microliters in 15mL or 1:150 dilution)<br />
# If OD is in linear range, calculate dilution for desired quantity of bacteria in 1mL (typically 10^8 cells /mL of chamber media)<br />
# Repeat dilution if not in linear range (using different ratio)<br />
# Inject 1mL of bacteria into each chamber (see below)<br />
<br />
===Fuel Cells===<br />
<br />
Procedure<br />
<br />
# Pipet 75mL of NaPiPES solution into each side of fuel cells<br />
# Inject 1mL of Resazurin solution into each side of chamber<br />
# Using Luer Lock nozzles, connect tubing from top ports to beaker w/ distilled water<br />
# Cut tubing to span distance from manifolds to each fuel cell (1 from nitrogen, 1 from air)<br />
# Attach syringe needles to tubing via Luer Lock nozzles<br />
# Using Luer Lock nozzles, connect tubing from manifolds<br />
# Start gas flow<br />
# Poke needles through bottom ports on fuel cells<br />
<br />
===Measurements===<br />
<br />
Procedure<br />
<br />
# Turn on computer and digital multimeter<br />
# Open LabVIEW program<br />
# Click Run arrow to take resistance readings<br />
# Connect fuel cells to resistor array via alligator clips<br />
# Click "Begin Current Readings" Icon on instrument display<br />
<br />
===Injections/Variables===<br />
<br />
Procedure<br />
<br />
# Once current readings reach equilibrium, inject bacteria into fuels cells using syringes<br />
# Allow bacteria to consume any carbon sources left in their media (approximately 12 hours)<br />
# Once current levels reach stable baseline, inject 1ml lactate solution<br />
# Inject additional variables as desired<br />
<br />
==Clean Up==<br />
<br />
Procedure<br />
<br />
# Drain chambers and soak in 70% ethanol<br />
# Remove carbon felt from anodes and discard (save titanium)<br />
# Scrub all parts in ethanol and distilled water successively<br />
# Use pipe cleaners on ports and tubes<br />
<br />
<br />
<br />
|}<br />
<br><br><br />
<br />
<!--- end body ---><br />
|}</div>Joyyhttp://2008.igem.org/Team:Harvard/Hardware/MFCProcedureTeam:Harvard/Hardware/MFCProcedure2008-10-30T04:04:02Z<p>Joyy: /* Constructing Fuel Cell Components */</p>
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|<br />
=Running an MFC Experiment=<br />
<br />
This page is intended as a comprehensive guide to completing a microbial fuel cell setup and running an experiment from start to finish. <br />
<br />
==Creating a Testing Environment==<br />
<br />
Begin 1-2 weeks prior to experiment<br />
<br />
===Constructing Fuel Cell Components===<br />
<br />
'''Materials (per fuel cell)'''<br />
<br />
* 4" Polycarbonate Square Tube, 2" Outer Diameter<br />
* 6" x 6" Polycarbonate Sheet, 1/4" Thick<br />
* 4 Steel Fully Threaded Stud, 1/4"-20 Thread, 6" Length<br />
* 8 Zinc Alloy Wing Flange Nut, 1/4"-20 Screw Size, 1" Wing Spread<br />
* 1" x 1" Nafion® membrane, 0.180mm thick<br />
* 1" x 1" Carbon felt, 0.25" thick<br />
* 1.5" x 1.5" E-TEK ELAT™ GDE (platinum on carbon)<br />
* 2' Titanium Grade 2 Wire .046" Diameter<br />
* Teflon Tape, 1/4" Width<br />
* 5" x 2.5" Silicone Sheet<br />
* Silicone Glue<br />
* Spiral Point Tap 1/4"-28<br />
* 8 Plastic Luer Lock Coupling Nylon, Female to Male Thread, 1/4"-28<br />
* 8 Luer Lock Injection Ports<br />
<br />
'''Procedure'''<br />
<br />
1) Mill Polycarbonate<br />
* Cut polycarbonate sheet into 4 equal 3" x 3" pieces<br />
* Drill four 3/8" holes through each piece, 1 per corner, indented 5mm from both sides<br />
[[Image:endplates.jpg | 600px]]<br />
* Cut polycarbonate tube into two equal 2" halves<br />
* Drill four 1/4" holes through each half in configuration shown<br />
[[Image:drilled_tube.jpg | 600px]]<br />
* Tap each hole with 1/4" -28 spiral tap<br />
<br />
2) Glue Chambers (repeat for each half)<br />
* Center tube on endplate by marking plate with 'X' from corner to corner<br />
* Squirt 2mm thick line of silicone on edge of tube (edge furthest from holes)<br />
* Press tube firmly against marked location on endplate<br />
* Quickly spread excess silicone along edge<br />
* Let stand 24h to harden<br />
[[Image:glued_half.jpg | 600px]]<br />
<br />
3) Construct Gaskets<br />
* Cut silicone sheet into two equal 2.25" x 2.25" pieces<br />
* Cut out centered inner squares in each piece, 1.75" x 1.75"<br />
* Using inner squares, cut two 'O' rings, inner diameter 1/4", outer diameter 1/2"<br />
[[Image:gaskets.jpg | 600px]]<br />
<br />
4) Construct Electrodes<br />
* Cut titanium wire into one 8" piece and one 16" piece<br />
* Using pliers, shape anode and cathode as shown<br />
'''Anode Frame'''<br><br />
[[Image:Anodeframe.jpg | 500px]]<br><br />
'''Cathode Frame'''<br><br />
[[Image:Cathodeframe.jpg | 500px]]<br />
<br />
* Spear carbon felt with tip of anode titanium wire and wedge into frame<br />
* Weave platinum carbon cloth through cathode titanium wire<br />
'''Anode'''<br><br />
[[Image:Anode_electrode.jpg | 500px]]<br><br />
'''Cathode'''<br><br />
[[Image:Cathode_electrode.jpg | 500px]]<br />
<br />
5) Seal Injection Ports<br />
* Wrap threads of all eight Luer Lock screws with 1' of teflon tape in opposite direction of screwing<br />
* Screw Luer Locks into all tapped holes in both chambers<br />
[[Image:fin_chamber.jpg | 600px]]<br />
<br />
===Setup of Measurement Device===<br />
<br />
'''Materials'''<br />
* Keithley 2700 Digital Multimeter<br />
* Keithley 7700 Multiplexer<br />
* Small Breadboard<br />
* Supply of insulted thin copper wire<br />
* 470 Ohm resistors (one/fuel cell)<br />
<br />
'''Procedure'''<br />
1) Wire Multiplexer<br />
* Open multiplexer, note channels<br />
* Cut two wire 18" wire leads per fuel cell<br />
* Strip ends, place one wire in each screw terminal, screw tight<br />
* Tape paired wires (two are attached to each channel) near non-attached ends and label<br />
* Clamp wire bundles near back of device with provided plastic latch clamps<br />
* Close Multiplexer and slide into 2700 DMM<br />
<br />
2) Create Resistor Array<br />
* Connect resistors across middle of breadboard (one per fuel cell)<br />
* Connect leads from multiplexer across resistors (one pair across each resistor)<br />
<br />
<br />
===Controlling the DMM with LabVIEW™===<br />
<br />
# Initialize Multimeter<br />
#* Attach 2700 to COM1 port of desktop computer w/ LabVIEW™<br />
#* Download our LabVIEW™ source code [[Media:MFCs.txt|MFCs.vi]]<br />
#* Open Program in LabVIEW™, adjust block diagram as necessary<br />
<br />
==Experiment Preparation==<br />
<br />
Begin 1 day prior to experiment<br />
<br />
===Assembling Chambers===<br />
<br />
'''Procedure'''<br />
1) Prepare Electrodes<br />
* Attach Luer Lock injection ports to all chamber screws<br />
* Poke tip of electrodes through designated ports from the inside<br />
[[Image:chamber_w_elec.jpg | 600px]]<br />
<br />
2 Align Chambers<br />
* Lay one chamber on a flat surface<br />
* Place silicone square ring on top edge of tube<br />
* Place polycarbonate square on silicone<br />
* Place silicone 'O' ring around central pore<br />
* Place Nafion membrane on top of 'O' ring<br />
* Sandwich membrane between second 'O' ring<br />
* Align second polycarbonate square on top of 'O' ring<br />
* Center second silicone square ring on polycarbonate square<br />
* Set second chamber on top of silicone, ensuring ports facing same direction as first chamber<br />
<br />
3) Clamp Chambers<br />
* Move assembly into vice or clamp<br />
* Insert rods through holes in end plates and screw on wing nuts<br />
* Tighten evenly<br />
<br />
===Solutions Prep===<br />
<br />
# Chamber media (150ml / fuel cell)<br />
#* 5.844 g/L 100mM NaCl<br />
#* 15.1185 g/L 50mM PIPES (hydrogen) <br />
#* ''7.0 pH''<br />
<br />
# Phosphate buffer (60ml / fuel cell)<br />
#* 2.918 g/L Monosodium phosphate, monohydrate<br />
#* 4.095 g/L Disodium phosphate, anhydrous <br />
#* 5.844 g/L 100mM NaCl<br />
#* ''7.0 pH''<br />
<br />
===Gas Tubing Assembly===<br />
<br />
Materials<br />
* 25' Silicone Soft Rubber Tubing, 3/32" ID, 7/32" OD, 1/16" Wall<br />
* Tank of Compressed Nitrogen<br />
* Gas Regulator<br />
* Lab Supply of Air<br />
* 4 Plastic Luer Lock Coupling Nylon, Male X Barb, for 3/32" Tube<br />
* 4 Plastic Luer Lock Coupling Nylon, Female X Barb, for 3/32" Tube<br />
* Plastic Luer Lock Coupling Nylon, T junctions, for 3/32" Tube<br />
* Syringe needles - 27 gauge<br />
* 2 Aspirator Flasks<br />
* 2 Rubber Stoppers<br />
<br />
Procedure<br />
# Make Flow Regulators<br />
#* Insert nozzle of female Luer Lock into rubber stoppers (poke hole if necessary)<br />
#* Cap aspirator flasks with rubber stoppers<br />
#* Attach tubing from gas sources to each glass nozzle of aspirator flask<br />
[[Image:flow_regs.jpg | 600px]]<br />
<br />
# Make Manifolds<br />
#* Attach T-junction Luer Lock pieces into manifold (1 junction/ fuel cell ; 2 manifods total)<br />
#* Turn last juction such that off is facing end of manifold<br />
#* Attach tubing from stopper of flow regulators to beginning of each mainfold<br />
[[Image:manifolds.jpg | 600px]]<br />
<br />
===Growing Strains===<br />
<br />
Materials<br />
<br />
* 150mL LB / strain<br />
* 1000mL airating flask / strain<br />
* Antibiotics (if desired strain must be selected for)<br />
* Plate or Glycerol Stock with desired strain<br />
<br />
Procedure<br />
<br />
# Fill flasks with LB<br />
# Add correct concentration of selection antibiotic<br />
# Pick single colony from plate, add to flask<br />
# Incubate overnight in shaker at 30C<br />
<br />
==Runtime==<br />
<br />
Begin 2 hours prior to experiment<br />
<br />
===Bacteria===<br />
<br />
Procedure<br />
<br />
# Pipet the cells out of the flask and into 250mL centrifuge containers <br />
# Make sure the containers are close in weight (within 0.5g of each other)<br />
# For a culture more than 250mL split it into two containers <br />
# Set the centrifuge temperature to 22-23C, spin speed to 5000RPM, and time to 15 min<br />
# After first spin, drain each container of the LB, making sure to leave the bacteria pellet intact<br />
# Resuspend pellet in 50mL of potassium buffer<br />
# After bacteria are fully resuspended (no pellets at all), spin down again, 22-23C, 5000RPM, 15min.<br />
# Pour potassium buffer out of container slowly.<br />
# Resuspend pellet in 50mL of potassium buffer<br />
# Spin down again, 22-23C, 5000RPM, 15min.<br />
# Pour potassium buffer out of container slowly. <br />
# Resuspend pellet in 4 mL of sodium pipes<br />
# Check OD (100microliters in 15mL or 1:150 dilution)<br />
# If OD is in linear range, calculate dilution for desired quantity of bacteria in 1mL (typically 10^8 cells /mL of chamber media)<br />
# Repeat dilution if not in linear range (using different ratio)<br />
# Inject 1mL of bacteria into each chamber (see below)<br />
<br />
===Fuel Cells===<br />
<br />
Procedure<br />
<br />
# Pipet 75mL of NaPiPES solution into each side of fuel cells<br />
# Inject 1mL of Resazurin solution into each side of chamber<br />
# Using Luer Lock nozzles, connect tubing from top ports to beaker w/ distilled water<br />
# Cut tubing to span distance from manifolds to each fuel cell (1 from nitrogen, 1 from air)<br />
# Attach syringe needles to tubing via Luer Lock nozzles<br />
# Using Luer Lock nozzles, connect tubing from manifolds<br />
# Start gas flow<br />
# Poke needles through bottom ports on fuel cells<br />
<br />
===Measurements===<br />
<br />
Procedure<br />
<br />
# Turn on computer and digital multimeter<br />
# Open LabVIEW program<br />
# Click Run arrow to take resistance readings<br />
# Connect fuel cells to resistor array via alligator clips<br />
# Click "Begin Current Readings" Icon on instrument display<br />
<br />
===Injections/Variables===<br />
<br />
Procedure<br />
<br />
# Once current readings reach equilibrium, inject bacteria into fuels cells using syringes<br />
# Allow bacteria to consume any carbon sources left in their media (approximately 12 hours)<br />
# Once current levels reach stable baseline, inject 1ml lactate solution<br />
# Inject additional variables as desired<br />
<br />
==Clean Up==<br />
<br />
Procedure<br />
<br />
# Drain chambers and soak in 70% ethanol<br />
# Remove carbon felt from anodes and discard (save titanium)<br />
# Scrub all parts in ethanol and distilled water successively<br />
# Use pipe cleaners on ports and tubes<br />
<br />
<br />
<br />
|}<br />
<br><br><br />
<br />
<!--- end body ---><br />
|}</div>Joyyhttp://2008.igem.org/Team:Harvard/Hardware/MFCProcedureTeam:Harvard/Hardware/MFCProcedure2008-10-30T04:03:18Z<p>Joyy: </p>
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{|align="justify" style="background-color:#FFFFFF;text-indent: 15pt;text-align:justify" cellpadding="50" width="90%"<br />
|<br />
=Running an MFC Experiment=<br />
<br />
This page is intended as a comprehensive guide to completing a microbial fuel cell setup and running an experiment from start to finish. <br />
<br />
==Creating a Testing Environment==<br />
<br />
Begin 1-2 weeks prior to experiment<br />
<br />
===Constructing Fuel Cell Components===<br />
<br />
'''Materials (per fuel cell)'''<br />
<br />
* 4" Polycarbonate Square Tube, 2" Outer Diameter<br />
* 6" x 6" Polycarbonate Sheet, 1/4" Thick<br />
* 4 Steel Fully Threaded Stud, 1/4"-20 Thread, 6" Length<br />
* 8 Zinc Alloy Wing Flange Nut, 1/4"-20 Screw Size, 1" Wing Spread<br />
* 1" x 1" Nafion® membrane, 0.180mm thick<br />
* 1" x 1" Carbon felt, 0.25" thick<br />
* 1.5" x 1.5" E-TEK ELAT™ GDE (platinum on carbon)<br />
* 2' Titanium Grade 2 Wire .046" Diameter<br />
* Teflon Tape, 1/4" Width<br />
* 5" x 2.5" Silicone Sheet<br />
* Silicone Glue<br />
* Spiral Point Tap 1/4"-28<br />
* 8 Plastic Luer Lock Coupling Nylon, Female to Male Thread, 1/4"-28<br />
* 8 Luer Lock Injection Ports<br />
<br />
'''Procedure'''<br />
<br />
*1) Mill Polycarbonate<br />
** Cut polycarbonate sheet into 4 equal 3" x 3" pieces<br />
** Drill four 3/8" holes through each piece, 1 per corner, indented 5mm from both sides<br />
[[Image:endplates.jpg | 600px]]<br />
** Cut polycarbonate tube into two equal 2" halves<br />
** Drill four 1/4" holes through each half in configuration shown<br />
[[Image:drilled_tube.jpg | 600px]]<br />
** Tap each hole with 1/4" -28 spiral tap<br />
<br />
*2) Glue Chambers (repeat for each half)<br />
** Center tube on endplate by marking plate with 'X' from corner to corner<br />
** Squirt 2mm thick line of silicone on edge of tube (edge furthest from holes)<br />
** Press tube firmly against marked location on endplate<br />
** Quickly spread excess silicone along edge<br />
** Let stand 24h to harden<br />
[[Image:glued_half.jpg | 600px]]<br />
<br />
*3) Construct Gaskets<br />
** Cut silicone sheet into two equal 2.25" x 2.25" pieces<br />
** Cut out centered inner squares in each piece, 1.75" x 1.75"<br />
** Using inner squares, cut two 'O' rings, inner diameter 1/4", outer diameter 1/2"<br />
[[Image:gaskets.jpg | 600px]]<br />
<br />
*4) Construct Electrodes<br />
** Cut titanium wire into one 8" piece and one 16" piece<br />
** Using pliers, shape anode and cathode as shown<br />
'''Anode Frame'''<br><br />
[[Image:Anodeframe.jpg | 500px]]<br><br />
'''Cathode Frame'''<br><br />
[[Image:Cathodeframe.jpg | 500px]]<br />
<br />
** Spear carbon felt with tip of anode titanium wire and wedge into frame<br />
** Weave platinum carbon cloth through cathode titanium wire<br />
'''Anode'''<br><br />
[[Image:Anode_electrode.jpg | 500px]]<br><br />
'''Cathode'''<br><br />
[[Image:Cathode_electrode.jpg | 500px]]<br />
<br />
*5) Seal Injection Ports<br />
** Wrap threads of all eight Luer Lock screws with 1' of teflon tape in opposite direction of screwing<br />
** Screw Luer Locks into all tapped holes in both chambers<br />
[[Image:fin_chamber.jpg | 600px]]<br />
<br />
===Setup of Measurement Device===<br />
<br />
'''Materials'''<br />
* Keithley 2700 Digital Multimeter<br />
* Keithley 7700 Multiplexer<br />
* Small Breadboard<br />
* Supply of insulted thin copper wire<br />
* 470 Ohm resistors (one/fuel cell)<br />
<br />
'''Procedure'''<br />
1) Wire Multiplexer<br />
* Open multiplexer, note channels<br />
* Cut two wire 18" wire leads per fuel cell<br />
* Strip ends, place one wire in each screw terminal, screw tight<br />
* Tape paired wires (two are attached to each channel) near non-attached ends and label<br />
* Clamp wire bundles near back of device with provided plastic latch clamps<br />
* Close Multiplexer and slide into 2700 DMM<br />
<br />
2) Create Resistor Array<br />
* Connect resistors across middle of breadboard (one per fuel cell)<br />
* Connect leads from multiplexer across resistors (one pair across each resistor)<br />
<br />
<br />
===Controlling the DMM with LabVIEW™===<br />
<br />
# Initialize Multimeter<br />
#* Attach 2700 to COM1 port of desktop computer w/ LabVIEW™<br />
#* Download our LabVIEW™ source code [[Media:MFCs.txt|MFCs.vi]]<br />
#* Open Program in LabVIEW™, adjust block diagram as necessary<br />
<br />
==Experiment Preparation==<br />
<br />
Begin 1 day prior to experiment<br />
<br />
===Assembling Chambers===<br />
<br />
'''Procedure'''<br />
1) Prepare Electrodes<br />
* Attach Luer Lock injection ports to all chamber screws<br />
* Poke tip of electrodes through designated ports from the inside<br />
[[Image:chamber_w_elec.jpg | 600px]]<br />
<br />
2 Align Chambers<br />
* Lay one chamber on a flat surface<br />
* Place silicone square ring on top edge of tube<br />
* Place polycarbonate square on silicone<br />
* Place silicone 'O' ring around central pore<br />
* Place Nafion membrane on top of 'O' ring<br />
* Sandwich membrane between second 'O' ring<br />
* Align second polycarbonate square on top of 'O' ring<br />
* Center second silicone square ring on polycarbonate square<br />
* Set second chamber on top of silicone, ensuring ports facing same direction as first chamber<br />
<br />
3) Clamp Chambers<br />
* Move assembly into vice or clamp<br />
* Insert rods through holes in end plates and screw on wing nuts<br />
* Tighten evenly<br />
<br />
===Solutions Prep===<br />
<br />
# Chamber media (150ml / fuel cell)<br />
#* 5.844 g/L 100mM NaCl<br />
#* 15.1185 g/L 50mM PIPES (hydrogen) <br />
#* ''7.0 pH''<br />
<br />
# Phosphate buffer (60ml / fuel cell)<br />
#* 2.918 g/L Monosodium phosphate, monohydrate<br />
#* 4.095 g/L Disodium phosphate, anhydrous <br />
#* 5.844 g/L 100mM NaCl<br />
#* ''7.0 pH''<br />
<br />
===Gas Tubing Assembly===<br />
<br />
Materials<br />
* 25' Silicone Soft Rubber Tubing, 3/32" ID, 7/32" OD, 1/16" Wall<br />
* Tank of Compressed Nitrogen<br />
* Gas Regulator<br />
* Lab Supply of Air<br />
* 4 Plastic Luer Lock Coupling Nylon, Male X Barb, for 3/32" Tube<br />
* 4 Plastic Luer Lock Coupling Nylon, Female X Barb, for 3/32" Tube<br />
* Plastic Luer Lock Coupling Nylon, T junctions, for 3/32" Tube<br />
* Syringe needles - 27 gauge<br />
* 2 Aspirator Flasks<br />
* 2 Rubber Stoppers<br />
<br />
Procedure<br />
# Make Flow Regulators<br />
#* Insert nozzle of female Luer Lock into rubber stoppers (poke hole if necessary)<br />
#* Cap aspirator flasks with rubber stoppers<br />
#* Attach tubing from gas sources to each glass nozzle of aspirator flask<br />
[[Image:flow_regs.jpg | 600px]]<br />
<br />
# Make Manifolds<br />
#* Attach T-junction Luer Lock pieces into manifold (1 junction/ fuel cell ; 2 manifods total)<br />
#* Turn last juction such that off is facing end of manifold<br />
#* Attach tubing from stopper of flow regulators to beginning of each mainfold<br />
[[Image:manifolds.jpg | 600px]]<br />
<br />
===Growing Strains===<br />
<br />
Materials<br />
<br />
* 150mL LB / strain<br />
* 1000mL airating flask / strain<br />
* Antibiotics (if desired strain must be selected for)<br />
* Plate or Glycerol Stock with desired strain<br />
<br />
Procedure<br />
<br />
# Fill flasks with LB<br />
# Add correct concentration of selection antibiotic<br />
# Pick single colony from plate, add to flask<br />
# Incubate overnight in shaker at 30C<br />
<br />
==Runtime==<br />
<br />
Begin 2 hours prior to experiment<br />
<br />
===Bacteria===<br />
<br />
Procedure<br />
<br />
# Pipet the cells out of the flask and into 250mL centrifuge containers <br />
# Make sure the containers are close in weight (within 0.5g of each other)<br />
# For a culture more than 250mL split it into two containers <br />
# Set the centrifuge temperature to 22-23C, spin speed to 5000RPM, and time to 15 min<br />
# After first spin, drain each container of the LB, making sure to leave the bacteria pellet intact<br />
# Resuspend pellet in 50mL of potassium buffer<br />
# After bacteria are fully resuspended (no pellets at all), spin down again, 22-23C, 5000RPM, 15min.<br />
# Pour potassium buffer out of container slowly.<br />
# Resuspend pellet in 50mL of potassium buffer<br />
# Spin down again, 22-23C, 5000RPM, 15min.<br />
# Pour potassium buffer out of container slowly. <br />
# Resuspend pellet in 4 mL of sodium pipes<br />
# Check OD (100microliters in 15mL or 1:150 dilution)<br />
# If OD is in linear range, calculate dilution for desired quantity of bacteria in 1mL (typically 10^8 cells /mL of chamber media)<br />
# Repeat dilution if not in linear range (using different ratio)<br />
# Inject 1mL of bacteria into each chamber (see below)<br />
<br />
===Fuel Cells===<br />
<br />
Procedure<br />
<br />
# Pipet 75mL of NaPiPES solution into each side of fuel cells<br />
# Inject 1mL of Resazurin solution into each side of chamber<br />
# Using Luer Lock nozzles, connect tubing from top ports to beaker w/ distilled water<br />
# Cut tubing to span distance from manifolds to each fuel cell (1 from nitrogen, 1 from air)<br />
# Attach syringe needles to tubing via Luer Lock nozzles<br />
# Using Luer Lock nozzles, connect tubing from manifolds<br />
# Start gas flow<br />
# Poke needles through bottom ports on fuel cells<br />
<br />
===Measurements===<br />
<br />
Procedure<br />
<br />
# Turn on computer and digital multimeter<br />
# Open LabVIEW program<br />
# Click Run arrow to take resistance readings<br />
# Connect fuel cells to resistor array via alligator clips<br />
# Click "Begin Current Readings" Icon on instrument display<br />
<br />
===Injections/Variables===<br />
<br />
Procedure<br />
<br />
# Once current readings reach equilibrium, inject bacteria into fuels cells using syringes<br />
# Allow bacteria to consume any carbon sources left in their media (approximately 12 hours)<br />
# Once current levels reach stable baseline, inject 1ml lactate solution<br />
# Inject additional variables as desired<br />
<br />
==Clean Up==<br />
<br />
Procedure<br />
<br />
# Drain chambers and soak in 70% ethanol<br />
# Remove carbon felt from anodes and discard (save titanium)<br />
# Scrub all parts in ethanol and distilled water successively<br />
# Use pipe cleaners on ports and tubes<br />
<br />
<br />
<br />
|}<br />
<br><br><br />
<br />
<!--- end body ---><br />
|}</div>Joyyhttp://2008.igem.org/Team:Harvard/Hardware/MFCProcedureTeam:Harvard/Hardware/MFCProcedure2008-10-30T04:01:48Z<p>Joyy: </p>
<hr />
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<!--- body here---><br />
{|align="justify" style="background-color:#FFFFFF;text-indent: 15pt;text-align:justify" cellpadding="50" width="90%"<br />
|<br />
=Running an MFC Experiment=<br />
<br />
This page is intended as a comprehensive guide to completing a microbial fuel cell setup and running an experiment from start to finish. <br />
<br />
==Creating a Testing Environment==<br />
<br />
Begin 1-2 weeks prior to experiment<br />
<br />
===Constructing Fuel Cell Components===<br />
<br />
'''Materials (per fuel cell)'''<br />
<br />
* 4" Polycarbonate Square Tube, 2" Outer Diameter<br />
* 6" x 6" Polycarbonate Sheet, 1/4" Thick<br />
* 4 Steel Fully Threaded Stud, 1/4"-20 Thread, 6" Length<br />
* 8 Zinc Alloy Wing Flange Nut, 1/4"-20 Screw Size, 1" Wing Spread<br />
* 1" x 1" Nafion® membrane, 0.180mm thick<br />
* 1" x 1" Carbon felt, 0.25" thick<br />
* 1.5" x 1.5" E-TEK ELAT™ GDE (platinum on carbon)<br />
* 2' Titanium Grade 2 Wire .046" Diameter<br />
* Teflon Tape, 1/4" Width<br />
* 5" x 2.5" Silicone Sheet<br />
* Silicone Glue<br />
* Spiral Point Tap 1/4"-28<br />
* 8 Plastic Luer Lock Coupling Nylon, Female to Male Thread, 1/4"-28<br />
* 8 Luer Lock Injection Ports<br />
<br />
'''Procedure'''<br />
<br />
*1) Mill Polycarbonate<br />
** Cut polycarbonate sheet into 4 equal 3" x 3" pieces<br />
** Drill four 3/8" holes through each piece, 1 per corner, indented 5mm from both sides<br />
[[Image:endplates.jpg | 600px]]<br />
** Cut polycarbonate tube into two equal 2" halves<br />
** Drill four 1/4" holes through each half in configuration shown<br />
[[Image:drilled_tube.jpg | 600px]]<br />
** Tap each hole with 1/4" -28 spiral tap<br />
<br />
*2) Glue Chambers (repeat for each half)<br />
** Center tube on endplate by marking plate with 'X' from corner to corner<br />
** Squirt 2mm thick line of silicone on edge of tube (edge furthest from holes)<br />
** Press tube firmly against marked location on endplate<br />
** Quickly spread excess silicone along edge<br />
** Let stand 24h to harden<br />
[[Image:glued_half.jpg | 600px]]<br />
<br />
*3) Construct Gaskets<br />
** Cut silicone sheet into two equal 2.25" x 2.25" pieces<br />
** Cut out centered inner squares in each piece, 1.75" x 1.75"<br />
** Using inner squares, cut two 'O' rings, inner diameter 1/4", outer diameter 1/2"<br />
[[Image:gaskets.jpg | 600px]]<br />
<br />
*4) Construct Electrodes<br />
** Cut titanium wire into one 8" piece and one 16" piece<br />
** Using pliers, shape anode and cathode as shown<br />
'''Anode Frame'''<br><br />
[[Image:Anodeframe.jpg | 500px]]<br><br />
'''Cathode Frame'''<br><br />
[[Image:Cathodeframe.jpg | 500px]]<br />
<br />
** Spear carbon felt with tip of anode titanium wire and wedge into frame<br />
** Weave platinum carbon cloth through cathode titanium wire<br />
'''Anode'''<br><br />
[[Image:Anode_electrode.jpg | 500px]]<br><br />
'''Cathode'''<br><br />
[[Image:Cathode_electrode.jpg | 500px]]<br />
<br />
*5) Seal Injection Ports<br />
** Wrap threads of all eight Luer Lock screws with 1' of teflon tape in opposite direction of screwing<br />
** Screw Luer Locks into all tapped holes in both chambers<br />
[[Image:fin_chamber.jpg | 600px]]<br />
<br />
===Setup of Measurement Device===<br />
<br />
'''Materials'''<br />
* Keithley 2700 Digital Multimeter<br />
* Keithley 7700 Multiplexer<br />
* Small Breadboard<br />
* Supply of insulted thin copper wire<br />
* 470 Ohm resistors (one/fuel cell)<br />
<br />
'''Procedure'''<br />
# Wire Multiplexer<br />
#* Open multiplexer, note channels<br />
#* Cut two wire 18" wire leads per fuel cell<br />
#* Strip ends, place one wire in each screw terminal, screw tight<br />
#* Tape paired wires (two are attached to each channel) near non-attached ends and label<br />
#* Clamp wire bundles near back of device with provided plastic latch clamps<br />
#* Close Multiplexer and slide into 2700 DMM<br />
<br />
# Create Resistor Array<br />
#* Connect resistors across middle of breadboard (one per fuel cell)<br />
#* Connect leads from multiplexer across resistors (one pair across each resistor)<br />
<br />
<br />
===Controlling the DMM with LabVIEW™===<br />
<br />
# Initialize Multimeter<br />
#* Attach 2700 to COM1 port of desktop computer w/ LabVIEW™<br />
#* Download our LabVIEW™ source code [[Media:MFCs.txt|MFCs.vi]]<br />
#* Open Program in LabVIEW™, adjust block diagram as necessary<br />
<br />
==Experiment Preparation==<br />
<br />
Begin 1 day prior to experiment<br />
<br />
===Assembling Chambers===<br />
<br />
Procedure<br />
# Prepare Electrodes<br />
#* Attach Luer Lock injection ports to all chamber screws<br />
#* Poke tip of electrodes through designated ports from the inside<br />
[[Image:chamber_w_elec.jpg | 600px]]<br />
<br />
# Align Chambers<br />
#* Lay one chamber on a flat surface<br />
#* Place silicone square ring on top edge of tube<br />
#* Place polycarbonate square on silicone<br />
#* Place silicone 'O' ring around central pore<br />
#* Place Nafion membrane on top of 'O' ring<br />
#* Sandwich membrane between second 'O' ring<br />
#* Align second polycarbonate square on top of 'O' ring<br />
#* Center second silicone square ring on polycarbonate square<br />
#* Set second chamber on top of silicone, ensuring ports facing same direction as first chamber<br />
<br />
# Clamp Chambers<br />
#* Move assembly into vice or clamp<br />
#* Insert rods through holes in end plates and screw on wing nuts<br />
#* Tighten evenly<br />
<br />
===Solutions Prep===<br />
<br />
# Chamber media (150ml / fuel cell)<br />
#* 5.844 g/L 100mM NaCl<br />
#* 15.1185 g/L 50mM PIPES (hydrogen) <br />
#* ''7.0 pH''<br />
<br />
# Phosphate buffer (60ml / fuel cell)<br />
#* 2.918 g/L Monosodium phosphate, monohydrate<br />
#* 4.095 g/L Disodium phosphate, anhydrous <br />
#* 5.844 g/L 100mM NaCl<br />
#* ''7.0 pH''<br />
<br />
===Gas Tubing Assembly===<br />
<br />
Materials<br />
* 25' Silicone Soft Rubber Tubing, 3/32" ID, 7/32" OD, 1/16" Wall<br />
* Tank of Compressed Nitrogen<br />
* Gas Regulator<br />
* Lab Supply of Air<br />
* 4 Plastic Luer Lock Coupling Nylon, Male X Barb, for 3/32" Tube<br />
* 4 Plastic Luer Lock Coupling Nylon, Female X Barb, for 3/32" Tube<br />
* Plastic Luer Lock Coupling Nylon, T junctions, for 3/32" Tube<br />
* Syringe needles - 27 gauge<br />
* 2 Aspirator Flasks<br />
* 2 Rubber Stoppers<br />
<br />
Procedure<br />
# Make Flow Regulators<br />
#* Insert nozzle of female Luer Lock into rubber stoppers (poke hole if necessary)<br />
#* Cap aspirator flasks with rubber stoppers<br />
#* Attach tubing from gas sources to each glass nozzle of aspirator flask<br />
[[Image:flow_regs.jpg | 600px]]<br />
<br />
# Make Manifolds<br />
#* Attach T-junction Luer Lock pieces into manifold (1 junction/ fuel cell ; 2 manifods total)<br />
#* Turn last juction such that off is facing end of manifold<br />
#* Attach tubing from stopper of flow regulators to beginning of each mainfold<br />
[[Image:manifolds.jpg | 600px]]<br />
<br />
===Growing Strains===<br />
<br />
Materials<br />
<br />
* 150mL LB / strain<br />
* 1000mL airating flask / strain<br />
* Antibiotics (if desired strain must be selected for)<br />
* Plate or Glycerol Stock with desired strain<br />
<br />
Procedure<br />
<br />
# Fill flasks with LB<br />
# Add correct concentration of selection antibiotic<br />
# Pick single colony from plate, add to flask<br />
# Incubate overnight in shaker at 30C<br />
<br />
==Runtime==<br />
<br />
Begin 2 hours prior to experiment<br />
<br />
===Bacteria===<br />
<br />
Procedure<br />
<br />
# Pipet the cells out of the flask and into 250mL centrifuge containers <br />
# Make sure the containers are close in weight (within 0.5g of each other)<br />
# For a culture more than 250mL split it into two containers <br />
# Set the centrifuge temperature to 22-23C, spin speed to 5000RPM, and time to 15 min<br />
# After first spin, drain each container of the LB, making sure to leave the bacteria pellet intact<br />
# Resuspend pellet in 50mL of potassium buffer<br />
# After bacteria are fully resuspended (no pellets at all), spin down again, 22-23C, 5000RPM, 15min.<br />
# Pour potassium buffer out of container slowly.<br />
# Resuspend pellet in 50mL of potassium buffer<br />
# Spin down again, 22-23C, 5000RPM, 15min.<br />
# Pour potassium buffer out of container slowly. <br />
# Resuspend pellet in 4 mL of sodium pipes<br />
# Check OD (100microliters in 15mL or 1:150 dilution)<br />
# If OD is in linear range, calculate dilution for desired quantity of bacteria in 1mL (typically 10^8 cells /mL of chamber media)<br />
# Repeat dilution if not in linear range (using different ratio)<br />
# Inject 1mL of bacteria into each chamber (see below)<br />
<br />
===Fuel Cells===<br />
<br />
Procedure<br />
<br />
# Pipet 75mL of NaPiPES solution into each side of fuel cells<br />
# Inject 1mL of Resazurin solution into each side of chamber<br />
# Using Luer Lock nozzles, connect tubing from top ports to beaker w/ distilled water<br />
# Cut tubing to span distance from manifolds to each fuel cell (1 from nitrogen, 1 from air)<br />
# Attach syringe needles to tubing via Luer Lock nozzles<br />
# Using Luer Lock nozzles, connect tubing from manifolds<br />
# Start gas flow<br />
# Poke needles through bottom ports on fuel cells<br />
<br />
===Measurements===<br />
<br />
Procedure<br />
<br />
# Turn on computer and digital multimeter<br />
# Open LabVIEW program<br />
# Click Run arrow to take resistance readings<br />
# Connect fuel cells to resistor array via alligator clips<br />
# Click "Begin Current Readings" Icon on instrument display<br />
<br />
===Injections/Variables===<br />
<br />
Procedure<br />
<br />
# Once current readings reach equilibrium, inject bacteria into fuels cells using syringes<br />
# Allow bacteria to consume any carbon sources left in their media (approximately 12 hours)<br />
# Once current levels reach stable baseline, inject 1ml lactate solution<br />
# Inject additional variables as desired<br />
<br />
==Clean Up==<br />
<br />
Procedure<br />
<br />
# Drain chambers and soak in 70% ethanol<br />
# Remove carbon felt from anodes and discard (save titanium)<br />
# Scrub all parts in ethanol and distilled water successively<br />
# Use pipe cleaners on ports and tubes<br />
<br />
<br />
<br />
|}<br />
<br><br><br />
<br />
<!--- end body ---><br />
|}</div>Joyyhttp://2008.igem.org/File:Manifolds.jpgFile:Manifolds.jpg2008-10-30T03:57:14Z<p>Joyy: </p>
<hr />
<div></div>Joyyhttp://2008.igem.org/File:Flow_regs.jpgFile:Flow regs.jpg2008-10-30T03:56:59Z<p>Joyy: </p>
<hr />
<div></div>Joyyhttp://2008.igem.org/File:Fin_chamber.jpgFile:Fin chamber.jpg2008-10-30T03:56:40Z<p>Joyy: </p>
<hr />
<div></div>Joyyhttp://2008.igem.org/Team:Harvard/Hardware/MFCProcedureTeam:Harvard/Hardware/MFCProcedure2008-10-30T03:56:28Z<p>Joyy: /* Assembling Chambers */</p>
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{|align="justify" style="background-color:#FFFFFF;text-indent: 15pt;text-align:justify" cellpadding="50" width="90%"<br />
|<br />
=Running an MFC Experiment=<br />
<br />
This page is intended as a comprehensive guide to completing a microbial fuel cell setup and running an experiment from start to finish. <br />
<br />
==Creating a Testing Environment==<br />
<br />
Begin 1-2 weeks prior to experiment<br />
<br />
===Constructing Fuel Cell Components===<br />
<br />
Materials (per fuel cell)<br />
<br />
* 4" Polycarbonate Square Tube, 2" Outer Diameter<br />
* 6" x 6" Polycarbonate Sheet, 1/4" Thick<br />
* 4 Steel Fully Threaded Stud, 1/4"-20 Thread, 6" Length<br />
* 8 Zinc Alloy Wing Flange Nut, 1/4"-20 Screw Size, 1" Wing Spread<br />
* 1" x 1" Nafion® membrane, 0.180mm thick<br />
* 1" x 1" Carbon felt, 0.25" thick<br />
* 1.5" x 1.5" E-TEK ELAT™ GDE (platinum on carbon)<br />
* 2' Titanium Grade 2 Wire .046" Diameter<br />
* Teflon Tape, 1/4" Width<br />
* 5" x 2.5" Silicone Sheet<br />
* Silicone Glue<br />
* Spiral Point Tap 1/4"-28<br />
* 8 Plastic Luer Lock Coupling Nylon, Female to Male Thread, 1/4"-28<br />
* 8 Luer Lock Injection Ports<br />
<br />
Procedure<br />
<br />
# Mill Polycarbonate<br />
#* Cut polycarbonate sheet into 4 equal 3" x 3" pieces<br />
#* Drill four 3/8" holes through each piece, 1 per corner, indented 5mm from both sides<br />
[[Image:endplates.jpg | 600px]]<br />
#* Cut polycarbonate tube into two equal 2" halves<br />
#* Drill four 1/4" holes through each half in configuration shown<br />
[[Image:drilled_tube.jpg | 600px]]<br />
#* Tap each hole with 1/4" -28 spiral tap<br />
<br />
# Glue Chambers (repeat for each half)<br />
#* Center tube on endplate by marking plate with 'X' from corner to corner<br />
#* Squirt 2mm thick line of silicone on edge of tube (edge furthest from holes)<br />
#* Press tube firmly against marked location on endplate<br />
#* Quickly spread excess silicone along edge<br />
#* Let stand 24h to harden<br />
[[Image:glued_half.jpg | 600px]]<br />
<br />
# Construct Gaskets<br />
#* Cut silicone sheet into two equal 2.25" x 2.25" pieces<br />
#* Cut out centered inner squares in each piece, 1.75" x 1.75"<br />
#* Using inner squares, cut two 'O' rings, inner diameter 1/4", outer diameter 1/2"<br />
[[Image:gaskets.jpg | 600px]]<br />
<br />
# Construct Electrodes<br />
#* Cut titanium wire into one 8" piece and one 16" piece<br />
#* Using pliers, shape anode and cathode as shown<br />
'''Anode Frame'''<br><br />
[[Image:Anodeframe.jpg | 500px]]<br><br />
'''Cathode Frame'''<br><br />
[[Image:Cathodeframe.jpg | 500px]]<br />
<br />
#* Spear carbon felt with tip of anode titanium wire and wedge into frame<br />
#* Weave platinum carbon cloth through cathode titanium wire<br />
'''Anode'''<br><br />
[[Image:Anode_electrode.jpg | 500px]]<br><br />
'''Cathode'''<br><br />
[[Image:Cathode_electrode.jpg | 500px]]<br />
<br />
# Seal Injection Ports<br />
#* Wrap threads of all eight Luer Lock screws with 1' of teflon tape in opposite direction of screwing<br />
#* Screw Luer Locks into all tapped holes in both chambers<br />
[[Image:fin_chamber.jpg | 600px]]<br />
<br />
===Setup of Measurement Device===<br />
<br />
Materials<br />
* Keithley 2700 Digital Multimeter<br />
* Keithley 7700 Multiplexer<br />
* Small Breadboard<br />
* Supply of insulted thin copper wire<br />
* 470 Ohm resistors (one/fuel cell)<br />
<br />
Procedure<br />
# Wire Multiplexer<br />
#* Open multiplexer, note channels<br />
#* Cut two wire 18" wire leads per fuel cell<br />
#* Strip ends, place one wire in each screw terminal, screw tight<br />
#* Tape paired wires (two are attached to each channel) near non-attached ends and label<br />
#* Clamp wire bundles near back of device with provided plastic latch clamps<br />
#* Close Multiplexer and slide into 2700 DMM<br />
<br />
# Create Resistor Array<br />
#* Connect resistors across middle of breadboard (one per fuel cell)<br />
#* Connect leads from multiplexer across resistors (one pair across each resistor)<br />
<br />
<br />
===Controlling the DMM with LabVIEW™===<br />
<br />
# Initialize Multimeter<br />
#* Attach 2700 to COM1 port of desktop computer w/ LabVIEW™<br />
#* Download our LabVIEW™ source code [[Media:MFCs.txt|MFCs.vi]]<br />
#* Open Program in LabVIEW™, adjust block diagram as necessary<br />
<br />
==Experiment Preparation==<br />
<br />
Begin 1 day prior to experiment<br />
<br />
===Assembling Chambers===<br />
<br />
Procedure<br />
# Prepare Electrodes<br />
#* Attach Luer Lock injection ports to all chamber screws<br />
#* Poke tip of electrodes through designated ports from the inside<br />
[[Image:chamber_w_elec.jpg | 600px]]<br />
<br />
# Align Chambers<br />
#* Lay one chamber on a flat surface<br />
#* Place silicone square ring on top edge of tube<br />
#* Place polycarbonate square on silicone<br />
#* Place silicone 'O' ring around central pore<br />
#* Place Nafion membrane on top of 'O' ring<br />
#* Sandwich membrane between second 'O' ring<br />
#* Align second polycarbonate square on top of 'O' ring<br />
#* Center second silicone square ring on polycarbonate square<br />
#* Set second chamber on top of silicone, ensuring ports facing same direction as first chamber<br />
<br />
# Clamp Chambers<br />
#* Move assembly into vice or clamp<br />
#* Insert rods through holes in end plates and screw on wing nuts<br />
#* Tighten evenly<br />
<br />
===Solutions Prep===<br />
<br />
# Chamber media (150ml / fuel cell)<br />
#* 5.844 g/L 100mM NaCl<br />
#* 15.1185 g/L 50mM PIPES (hydrogen) <br />
#* ''7.0 pH''<br />
<br />
# Phosphate buffer (60ml / fuel cell)<br />
#* 2.918 g/L Monosodium phosphate, monohydrate<br />
#* 4.095 g/L Disodium phosphate, anhydrous <br />
#* 5.844 g/L 100mM NaCl<br />
#* ''7.0 pH''<br />
<br />
===Gas Tubing Assembly===<br />
<br />
Materials<br />
* 25' Silicone Soft Rubber Tubing, 3/32" ID, 7/32" OD, 1/16" Wall<br />
* Tank of Compressed Nitrogen<br />
* Gas Regulator<br />
* Lab Supply of Air<br />
* 4 Plastic Luer Lock Coupling Nylon, Male X Barb, for 3/32" Tube<br />
* 4 Plastic Luer Lock Coupling Nylon, Female X Barb, for 3/32" Tube<br />
* Plastic Luer Lock Coupling Nylon, T junctions, for 3/32" Tube<br />
* Syringe needles - 27 gauge<br />
* 2 Aspirator Flasks<br />
* 2 Rubber Stoppers<br />
<br />
Procedure<br />
# Make Flow Regulators<br />
#* Insert nozzle of female Luer Lock into rubber stoppers (poke hole if necessary)<br />
#* Cap aspirator flasks with rubber stoppers<br />
#* Attach tubing from gas sources to each glass nozzle of aspirator flask<br />
[[Image:flow_regs.jpg | 600px]]<br />
<br />
# Make Manifolds<br />
#* Attach T-junction Luer Lock pieces into manifold (1 junction/ fuel cell ; 2 manifods total)<br />
#* Turn last juction such that off is facing end of manifold<br />
#* Attach tubing from stopper of flow regulators to beginning of each mainfold<br />
[[Image:manifolds.jpg | 600px]]<br />
<br />
===Growing Strains===<br />
<br />
Materials<br />
<br />
* 150mL LB / strain<br />
* 1000mL airating flask / strain<br />
* Antibiotics (if desired strain must be selected for)<br />
* Plate or Glycerol Stock with desired strain<br />
<br />
Procedure<br />
<br />
# Fill flasks with LB<br />
# Add correct concentration of selection antibiotic<br />
# Pick single colony from plate, add to flask<br />
# Incubate overnight in shaker at 30C<br />
<br />
==Runtime==<br />
<br />
Begin 2 hours prior to experiment<br />
<br />
===Bacteria===<br />
<br />
Procedure<br />
<br />
# Pipet the cells out of the flask and into 250mL centrifuge containers <br />
# Make sure the containers are close in weight (within 0.5g of each other)<br />
# For a culture more than 250mL split it into two containers <br />
# Set the centrifuge temperature to 22-23C, spin speed to 5000RPM, and time to 15 min<br />
# After first spin, drain each container of the LB, making sure to leave the bacteria pellet intact<br />
# Resuspend pellet in 50mL of potassium buffer<br />
# After bacteria are fully resuspended (no pellets at all), spin down again, 22-23C, 5000RPM, 15min.<br />
# Pour potassium buffer out of container slowly.<br />
# Resuspend pellet in 50mL of potassium buffer<br />
# Spin down again, 22-23C, 5000RPM, 15min.<br />
# Pour potassium buffer out of container slowly. <br />
# Resuspend pellet in 4 mL of sodium pipes<br />
# Check OD (100microliters in 15mL or 1:150 dilution)<br />
# If OD is in linear range, calculate dilution for desired quantity of bacteria in 1mL (typically 10^8 cells /mL of chamber media)<br />
# Repeat dilution if not in linear range (using different ratio)<br />
# Inject 1mL of bacteria into each chamber (see below)<br />
<br />
===Fuel Cells===<br />
<br />
Procedure<br />
<br />
# Pipet 75mL of NaPiPES solution into each side of fuel cells<br />
# Inject 1mL of Resazurin solution into each side of chamber<br />
# Using Luer Lock nozzles, connect tubing from top ports to beaker w/ distilled water<br />
# Cut tubing to span distance from manifolds to each fuel cell (1 from nitrogen, 1 from air)<br />
# Attach syringe needles to tubing via Luer Lock nozzles<br />
# Using Luer Lock nozzles, connect tubing from manifolds<br />
# Start gas flow<br />
# Poke needles through bottom ports on fuel cells<br />
<br />
===Measurements===<br />
<br />
Procedure<br />
<br />
# Turn on computer and digital multimeter<br />
# Open LabVIEW program<br />
# Click Run arrow to take resistance readings<br />
# Connect fuel cells to resistor array via alligator clips<br />
# Click "Begin Current Readings" Icon on instrument display<br />
<br />
===Injections/Variables===<br />
<br />
Procedure<br />
<br />
# Once current readings reach equilibrium, inject bacteria into fuels cells using syringes<br />
# Allow bacteria to consume any carbon sources left in their media (approximately 12 hours)<br />
# Once current levels reach stable baseline, inject 1ml lactate solution<br />
# Inject additional variables as desired<br />
<br />
==Clean Up==<br />
<br />
Procedure<br />
<br />
# Drain chambers and soak in 70% ethanol<br />
# Remove carbon felt from anodes and discard (save titanium)<br />
# Scrub all parts in ethanol and distilled water successively<br />
# Use pipe cleaners on ports and tubes<br />
<br />
<br />
<br />
|}<br />
<br><br><br />
<br />
<!--- end body ---><br />
|}</div>Joyyhttp://2008.igem.org/File:Chamber_w_elec.jpgFile:Chamber w elec.jpg2008-10-30T03:56:12Z<p>Joyy: </p>
<hr />
<div></div>Joyyhttp://2008.igem.org/File:Cathode_electrode.jpgFile:Cathode electrode.jpg2008-10-30T03:55:40Z<p>Joyy: </p>
<hr />
<div></div>Joyyhttp://2008.igem.org/File:Anode_electrode.jpgFile:Anode electrode.jpg2008-10-30T03:55:21Z<p>Joyy: </p>
<hr />
<div></div>Joyyhttp://2008.igem.org/File:Cathodeframe.jpgFile:Cathodeframe.jpg2008-10-30T03:54:59Z<p>Joyy: </p>
<hr />
<div></div>Joyyhttp://2008.igem.org/Team:Harvard/Hardware/MFCProcedureTeam:Harvard/Hardware/MFCProcedure2008-10-30T03:54:40Z<p>Joyy: /* Constructing Fuel Cell Components */</p>
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<!--- body here---><br />
{|align="justify" style="background-color:#FFFFFF;text-indent: 15pt;text-align:justify" cellpadding="50" width="90%"<br />
|<br />
=Running an MFC Experiment=<br />
<br />
This page is intended as a comprehensive guide to completing a microbial fuel cell setup and running an experiment from start to finish. <br />
<br />
==Creating a Testing Environment==<br />
<br />
Begin 1-2 weeks prior to experiment<br />
<br />
===Constructing Fuel Cell Components===<br />
<br />
Materials (per fuel cell)<br />
<br />
* 4" Polycarbonate Square Tube, 2" Outer Diameter<br />
* 6" x 6" Polycarbonate Sheet, 1/4" Thick<br />
* 4 Steel Fully Threaded Stud, 1/4"-20 Thread, 6" Length<br />
* 8 Zinc Alloy Wing Flange Nut, 1/4"-20 Screw Size, 1" Wing Spread<br />
* 1" x 1" Nafion® membrane, 0.180mm thick<br />
* 1" x 1" Carbon felt, 0.25" thick<br />
* 1.5" x 1.5" E-TEK ELAT™ GDE (platinum on carbon)<br />
* 2' Titanium Grade 2 Wire .046" Diameter<br />
* Teflon Tape, 1/4" Width<br />
* 5" x 2.5" Silicone Sheet<br />
* Silicone Glue<br />
* Spiral Point Tap 1/4"-28<br />
* 8 Plastic Luer Lock Coupling Nylon, Female to Male Thread, 1/4"-28<br />
* 8 Luer Lock Injection Ports<br />
<br />
Procedure<br />
<br />
# Mill Polycarbonate<br />
#* Cut polycarbonate sheet into 4 equal 3" x 3" pieces<br />
#* Drill four 3/8" holes through each piece, 1 per corner, indented 5mm from both sides<br />
[[Image:endplates.jpg | 600px]]<br />
#* Cut polycarbonate tube into two equal 2" halves<br />
#* Drill four 1/4" holes through each half in configuration shown<br />
[[Image:drilled_tube.jpg | 600px]]<br />
#* Tap each hole with 1/4" -28 spiral tap<br />
<br />
# Glue Chambers (repeat for each half)<br />
#* Center tube on endplate by marking plate with 'X' from corner to corner<br />
#* Squirt 2mm thick line of silicone on edge of tube (edge furthest from holes)<br />
#* Press tube firmly against marked location on endplate<br />
#* Quickly spread excess silicone along edge<br />
#* Let stand 24h to harden<br />
[[Image:glued_half.jpg | 600px]]<br />
<br />
# Construct Gaskets<br />
#* Cut silicone sheet into two equal 2.25" x 2.25" pieces<br />
#* Cut out centered inner squares in each piece, 1.75" x 1.75"<br />
#* Using inner squares, cut two 'O' rings, inner diameter 1/4", outer diameter 1/2"<br />
[[Image:gaskets.jpg | 600px]]<br />
<br />
# Construct Electrodes<br />
#* Cut titanium wire into one 8" piece and one 16" piece<br />
#* Using pliers, shape anode and cathode as shown<br />
'''Anode Frame'''<br><br />
[[Image:Anodeframe.jpg | 500px]]<br><br />
'''Cathode Frame'''<br><br />
[[Image:Cathodeframe.jpg | 500px]]<br />
<br />
#* Spear carbon felt with tip of anode titanium wire and wedge into frame<br />
#* Weave platinum carbon cloth through cathode titanium wire<br />
'''Anode'''<br><br />
[[Image:Anode_electrode.jpg | 500px]]<br><br />
'''Cathode'''<br><br />
[[Image:Cathode_electrode.jpg | 500px]]<br />
<br />
# Seal Injection Ports<br />
#* Wrap threads of all eight Luer Lock screws with 1' of teflon tape in opposite direction of screwing<br />
#* Screw Luer Locks into all tapped holes in both chambers<br />
[[Image:fin_chamber.jpg | 600px]]<br />
<br />
===Setup of Measurement Device===<br />
<br />
Materials<br />
* Keithley 2700 Digital Multimeter<br />
* Keithley 7700 Multiplexer<br />
* Small Breadboard<br />
* Supply of insulted thin copper wire<br />
* 470 Ohm resistors (one/fuel cell)<br />
<br />
Procedure<br />
# Wire Multiplexer<br />
#* Open multiplexer, note channels<br />
#* Cut two wire 18" wire leads per fuel cell<br />
#* Strip ends, place one wire in each screw terminal, screw tight<br />
#* Tape paired wires (two are attached to each channel) near non-attached ends and label<br />
#* Clamp wire bundles near back of device with provided plastic latch clamps<br />
#* Close Multiplexer and slide into 2700 DMM<br />
<br />
# Create Resistor Array<br />
#* Connect resistors across middle of breadboard (one per fuel cell)<br />
#* Connect leads from multiplexer across resistors (one pair across each resistor)<br />
<br />
<br />
===Controlling the DMM with LabVIEW™===<br />
<br />
# Initialize Multimeter<br />
#* Attach 2700 to COM1 port of desktop computer w/ LabVIEW™<br />
#* Download our LabVIEW™ source code [[Media:MFCs.txt|MFCs.vi]]<br />
#* Open Program in LabVIEW™, adjust block diagram as necessary<br />
<br />
==Experiment Preparation==<br />
<br />
Begin 1 day prior to experiment<br />
<br />
===Assembling Chambers===<br />
<br />
Procedure<br />
# Prepare Electrodes<br />
#* Attach Luer Lock injection ports to all chamber screws<br />
#* Poke tip of electrodes through designated ports from the inside<br />
[[Image:chamber_w_elec.jpg | 600px]]<br />
<br />
# Align Chambers<br />
#* Lay one chamber on a flat surface<br />
#* Place silicone square ring on top edge of tube<br />
#* Place polycarbonate square on silicone<br />
#* Place silicone 'O' ring around central pore<br />
#* Place Nafion membrane on top of 'O' ring<br />
#* Sandwich membrane between second 'O' ring<br />
#* Align second polycarbonate square on top of 'O' ring<br />
#* Center second silicone square ring on polycarbonate square<br />
#* Set second chamber on top of silicone, ensuring ports facing same direction as first chamber<br />
<br />
# Clamp Chambers<br />
#* Move assembly into vice or clamp<br />
#* Insert rods through holes in end plates and screw on wing nuts<br />
#* Tighten evenly<br />
[[Image:fuelcells.jpg | 600px]]<br />
<br />
===Solutions Prep===<br />
<br />
# Chamber media (150ml / fuel cell)<br />
#* 5.844 g/L 100mM NaCl<br />
#* 15.1185 g/L 50mM PIPES (hydrogen) <br />
#* ''7.0 pH''<br />
<br />
# Phosphate buffer (60ml / fuel cell)<br />
#* 2.918 g/L Monosodium phosphate, monohydrate<br />
#* 4.095 g/L Disodium phosphate, anhydrous <br />
#* 5.844 g/L 100mM NaCl<br />
#* ''7.0 pH''<br />
<br />
===Gas Tubing Assembly===<br />
<br />
Materials<br />
* 25' Silicone Soft Rubber Tubing, 3/32" ID, 7/32" OD, 1/16" Wall<br />
* Tank of Compressed Nitrogen<br />
* Gas Regulator<br />
* Lab Supply of Air<br />
* 4 Plastic Luer Lock Coupling Nylon, Male X Barb, for 3/32" Tube<br />
* 4 Plastic Luer Lock Coupling Nylon, Female X Barb, for 3/32" Tube<br />
* Plastic Luer Lock Coupling Nylon, T junctions, for 3/32" Tube<br />
* Syringe needles - 27 gauge<br />
* 2 Aspirator Flasks<br />
* 2 Rubber Stoppers<br />
<br />
Procedure<br />
# Make Flow Regulators<br />
#* Insert nozzle of female Luer Lock into rubber stoppers (poke hole if necessary)<br />
#* Cap aspirator flasks with rubber stoppers<br />
#* Attach tubing from gas sources to each glass nozzle of aspirator flask<br />
[[Image:flow_regs.jpg | 600px]]<br />
<br />
# Make Manifolds<br />
#* Attach T-junction Luer Lock pieces into manifold (1 junction/ fuel cell ; 2 manifods total)<br />
#* Turn last juction such that off is facing end of manifold<br />
#* Attach tubing from stopper of flow regulators to beginning of each mainfold<br />
[[Image:manifolds.jpg | 600px]]<br />
<br />
===Growing Strains===<br />
<br />
Materials<br />
<br />
* 150mL LB / strain<br />
* 1000mL airating flask / strain<br />
* Antibiotics (if desired strain must be selected for)<br />
* Plate or Glycerol Stock with desired strain<br />
<br />
Procedure<br />
<br />
# Fill flasks with LB<br />
# Add correct concentration of selection antibiotic<br />
# Pick single colony from plate, add to flask<br />
# Incubate overnight in shaker at 30C<br />
<br />
==Runtime==<br />
<br />
Begin 2 hours prior to experiment<br />
<br />
===Bacteria===<br />
<br />
Procedure<br />
<br />
# Pipet the cells out of the flask and into 250mL centrifuge containers <br />
# Make sure the containers are close in weight (within 0.5g of each other)<br />
# For a culture more than 250mL split it into two containers <br />
# Set the centrifuge temperature to 22-23C, spin speed to 5000RPM, and time to 15 min<br />
# After first spin, drain each container of the LB, making sure to leave the bacteria pellet intact<br />
# Resuspend pellet in 50mL of potassium buffer<br />
# After bacteria are fully resuspended (no pellets at all), spin down again, 22-23C, 5000RPM, 15min.<br />
# Pour potassium buffer out of container slowly.<br />
# Resuspend pellet in 50mL of potassium buffer<br />
# Spin down again, 22-23C, 5000RPM, 15min.<br />
# Pour potassium buffer out of container slowly. <br />
# Resuspend pellet in 4 mL of sodium pipes<br />
# Check OD (100microliters in 15mL or 1:150 dilution)<br />
# If OD is in linear range, calculate dilution for desired quantity of bacteria in 1mL (typically 10^8 cells /mL of chamber media)<br />
# Repeat dilution if not in linear range (using different ratio)<br />
# Inject 1mL of bacteria into each chamber (see below)<br />
<br />
===Fuel Cells===<br />
<br />
Procedure<br />
<br />
# Pipet 75mL of NaPiPES solution into each side of fuel cells<br />
# Inject 1mL of Resazurin solution into each side of chamber<br />
# Using Luer Lock nozzles, connect tubing from top ports to beaker w/ distilled water<br />
# Cut tubing to span distance from manifolds to each fuel cell (1 from nitrogen, 1 from air)<br />
# Attach syringe needles to tubing via Luer Lock nozzles<br />
# Using Luer Lock nozzles, connect tubing from manifolds<br />
# Start gas flow<br />
# Poke needles through bottom ports on fuel cells<br />
<br />
===Measurements===<br />
<br />
Procedure<br />
<br />
# Turn on computer and digital multimeter<br />
# Open LabVIEW program<br />
# Click Run arrow to take resistance readings<br />
# Connect fuel cells to resistor array via alligator clips<br />
# Click "Begin Current Readings" Icon on instrument display<br />
<br />
===Injections/Variables===<br />
<br />
Procedure<br />
<br />
# Once current readings reach equilibrium, inject bacteria into fuels cells using syringes<br />
# Allow bacteria to consume any carbon sources left in their media (approximately 12 hours)<br />
# Once current levels reach stable baseline, inject 1ml lactate solution<br />
# Inject additional variables as desired<br />
<br />
==Clean Up==<br />
<br />
Procedure<br />
<br />
# Drain chambers and soak in 70% ethanol<br />
# Remove carbon felt from anodes and discard (save titanium)<br />
# Scrub all parts in ethanol and distilled water successively<br />
# Use pipe cleaners on ports and tubes<br />
<br />
<br />
<br />
|}<br />
<br><br><br />
<br />
<!--- end body ---><br />
|}</div>Joyyhttp://2008.igem.org/Team:Harvard/Hardware/MFCProcedureTeam:Harvard/Hardware/MFCProcedure2008-10-30T03:53:42Z<p>Joyy: /* Constructing Fuel Cell Components */</p>
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|<br />
=Running an MFC Experiment=<br />
<br />
This page is intended as a comprehensive guide to completing a microbial fuel cell setup and running an experiment from start to finish. <br />
<br />
==Creating a Testing Environment==<br />
<br />
Begin 1-2 weeks prior to experiment<br />
<br />
===Constructing Fuel Cell Components===<br />
<br />
Materials (per fuel cell)<br />
<br />
* 4" Polycarbonate Square Tube, 2" Outer Diameter<br />
* 6" x 6" Polycarbonate Sheet, 1/4" Thick<br />
* 4 Steel Fully Threaded Stud, 1/4"-20 Thread, 6" Length<br />
* 8 Zinc Alloy Wing Flange Nut, 1/4"-20 Screw Size, 1" Wing Spread<br />
* 1" x 1" Nafion® membrane, 0.180mm thick<br />
* 1" x 1" Carbon felt, 0.25" thick<br />
* 1.5" x 1.5" E-TEK ELAT™ GDE (platinum on carbon)<br />
* 2' Titanium Grade 2 Wire .046" Diameter<br />
* Teflon Tape, 1/4" Width<br />
* 5" x 2.5" Silicone Sheet<br />
* Silicone Glue<br />
* Spiral Point Tap 1/4"-28<br />
* 8 Plastic Luer Lock Coupling Nylon, Female to Male Thread, 1/4"-28<br />
* 8 Luer Lock Injection Ports<br />
<br />
Procedure<br />
<br />
# Mill Polycarbonate<br />
#* Cut polycarbonate sheet into 4 equal 3" x 3" pieces<br />
#* Drill four 3/8" holes through each piece, 1 per corner, indented 5mm from both sides<br />
[[Image:endplates.jpg | 600px]]<br />
#* Cut polycarbonate tube into two equal 2" halves<br />
#* Drill four 1/4" holes through each half in configuration shown<br />
[[Image:drilled_tube.jpg | 600px]]<br />
#* Tap each hole with 1/4" -28 spiral tap<br />
<br />
# Glue Chambers (repeat for each half)<br />
#* Center tube on endplate by marking plate with 'X' from corner to corner<br />
#* Squirt 2mm thick line of silicone on edge of tube (edge furthest from holes)<br />
#* Press tube firmly against marked location on endplate<br />
#* Quickly spread excess silicone along edge<br />
#* Let stand 24h to harden<br />
[[Image:glued_half.jpg | 600px]]<br />
<br />
# Construct Gaskets<br />
#* Cut silicone sheet into two equal 2.25" x 2.25" pieces<br />
#* Cut out centered inner squares in each piece, 1.75" x 1.75"<br />
#* Using inner squares, cut two 'O' rings, inner diameter 1/4", outer diameter 1/2"<br />
[[Image:gaskets.jpg | 600px]]<br />
<br />
# Construct Electrodes<br />
#* Cut titanium wire into one 8" piece and one 16" piece<br />
#* Using pliers, shape anode and cathode as shown<br />
'''Anode Frame'''<br><br />
[[Image:Anodeframe.jpg | 300px]]<br><br />
'''Cathode Frame'''<br><br />
[[Image:Cathodeframe.jpg | 300px]]<br />
<br />
#* Spear carbon felt with tip of anode titanium wire and wedge into frame<br />
#* Weave platinum carbon cloth through cathode titanium wire<br />
'''Anode'''<br><br />
[[Image:Anode_electrode.jpg | 300px]]<br><br />
'''Cathode'''<br><br />
[[Image:Cathode_electrode.jpg | 300px]]<br />
<br />
# Seal Injection Ports<br />
#* Wrap threads of all eight Luer Lock screws with 1' of teflon tape in opposite direction of screwing<br />
#* Screw Luer Locks into all tapped holes in both chambers<br />
[[Image:fin_chamber.jpg | 600px]]<br />
<br />
===Setup of Measurement Device===<br />
<br />
Materials<br />
* Keithley 2700 Digital Multimeter<br />
* Keithley 7700 Multiplexer<br />
* Small Breadboard<br />
* Supply of insulted thin copper wire<br />
* 470 Ohm resistors (one/fuel cell)<br />
<br />
Procedure<br />
# Wire Multiplexer<br />
#* Open multiplexer, note channels<br />
#* Cut two wire 18" wire leads per fuel cell<br />
#* Strip ends, place one wire in each screw terminal, screw tight<br />
#* Tape paired wires (two are attached to each channel) near non-attached ends and label<br />
#* Clamp wire bundles near back of device with provided plastic latch clamps<br />
#* Close Multiplexer and slide into 2700 DMM<br />
<br />
# Create Resistor Array<br />
#* Connect resistors across middle of breadboard (one per fuel cell)<br />
#* Connect leads from multiplexer across resistors (one pair across each resistor)<br />
<br />
<br />
===Controlling the DMM with LabVIEW™===<br />
<br />
# Initialize Multimeter<br />
#* Attach 2700 to COM1 port of desktop computer w/ LabVIEW™<br />
#* Download our LabVIEW™ source code [[Media:MFCs.txt|MFCs.vi]]<br />
#* Open Program in LabVIEW™, adjust block diagram as necessary<br />
<br />
==Experiment Preparation==<br />
<br />
Begin 1 day prior to experiment<br />
<br />
===Assembling Chambers===<br />
<br />
Procedure<br />
# Prepare Electrodes<br />
#* Attach Luer Lock injection ports to all chamber screws<br />
#* Poke tip of electrodes through designated ports from the inside<br />
[[Image:chamber_w_elec.jpg | 600px]]<br />
<br />
# Align Chambers<br />
#* Lay one chamber on a flat surface<br />
#* Place silicone square ring on top edge of tube<br />
#* Place polycarbonate square on silicone<br />
#* Place silicone 'O' ring around central pore<br />
#* Place Nafion membrane on top of 'O' ring<br />
#* Sandwich membrane between second 'O' ring<br />
#* Align second polycarbonate square on top of 'O' ring<br />
#* Center second silicone square ring on polycarbonate square<br />
#* Set second chamber on top of silicone, ensuring ports facing same direction as first chamber<br />
<br />
# Clamp Chambers<br />
#* Move assembly into vice or clamp<br />
#* Insert rods through holes in end plates and screw on wing nuts<br />
#* Tighten evenly<br />
[[Image:fuelcells.jpg | 600px]]<br />
<br />
===Solutions Prep===<br />
<br />
# Chamber media (150ml / fuel cell)<br />
#* 5.844 g/L 100mM NaCl<br />
#* 15.1185 g/L 50mM PIPES (hydrogen) <br />
#* ''7.0 pH''<br />
<br />
# Phosphate buffer (60ml / fuel cell)<br />
#* 2.918 g/L Monosodium phosphate, monohydrate<br />
#* 4.095 g/L Disodium phosphate, anhydrous <br />
#* 5.844 g/L 100mM NaCl<br />
#* ''7.0 pH''<br />
<br />
===Gas Tubing Assembly===<br />
<br />
Materials<br />
* 25' Silicone Soft Rubber Tubing, 3/32" ID, 7/32" OD, 1/16" Wall<br />
* Tank of Compressed Nitrogen<br />
* Gas Regulator<br />
* Lab Supply of Air<br />
* 4 Plastic Luer Lock Coupling Nylon, Male X Barb, for 3/32" Tube<br />
* 4 Plastic Luer Lock Coupling Nylon, Female X Barb, for 3/32" Tube<br />
* Plastic Luer Lock Coupling Nylon, T junctions, for 3/32" Tube<br />
* Syringe needles - 27 gauge<br />
* 2 Aspirator Flasks<br />
* 2 Rubber Stoppers<br />
<br />
Procedure<br />
# Make Flow Regulators<br />
#* Insert nozzle of female Luer Lock into rubber stoppers (poke hole if necessary)<br />
#* Cap aspirator flasks with rubber stoppers<br />
#* Attach tubing from gas sources to each glass nozzle of aspirator flask<br />
[[Image:flow_regs.jpg | 600px]]<br />
<br />
# Make Manifolds<br />
#* Attach T-junction Luer Lock pieces into manifold (1 junction/ fuel cell ; 2 manifods total)<br />
#* Turn last juction such that off is facing end of manifold<br />
#* Attach tubing from stopper of flow regulators to beginning of each mainfold<br />
[[Image:manifolds.jpg | 600px]]<br />
<br />
===Growing Strains===<br />
<br />
Materials<br />
<br />
* 150mL LB / strain<br />
* 1000mL airating flask / strain<br />
* Antibiotics (if desired strain must be selected for)<br />
* Plate or Glycerol Stock with desired strain<br />
<br />
Procedure<br />
<br />
# Fill flasks with LB<br />
# Add correct concentration of selection antibiotic<br />
# Pick single colony from plate, add to flask<br />
# Incubate overnight in shaker at 30C<br />
<br />
==Runtime==<br />
<br />
Begin 2 hours prior to experiment<br />
<br />
===Bacteria===<br />
<br />
Procedure<br />
<br />
# Pipet the cells out of the flask and into 250mL centrifuge containers <br />
# Make sure the containers are close in weight (within 0.5g of each other)<br />
# For a culture more than 250mL split it into two containers <br />
# Set the centrifuge temperature to 22-23C, spin speed to 5000RPM, and time to 15 min<br />
# After first spin, drain each container of the LB, making sure to leave the bacteria pellet intact<br />
# Resuspend pellet in 50mL of potassium buffer<br />
# After bacteria are fully resuspended (no pellets at all), spin down again, 22-23C, 5000RPM, 15min.<br />
# Pour potassium buffer out of container slowly.<br />
# Resuspend pellet in 50mL of potassium buffer<br />
# Spin down again, 22-23C, 5000RPM, 15min.<br />
# Pour potassium buffer out of container slowly. <br />
# Resuspend pellet in 4 mL of sodium pipes<br />
# Check OD (100microliters in 15mL or 1:150 dilution)<br />
# If OD is in linear range, calculate dilution for desired quantity of bacteria in 1mL (typically 10^8 cells /mL of chamber media)<br />
# Repeat dilution if not in linear range (using different ratio)<br />
# Inject 1mL of bacteria into each chamber (see below)<br />
<br />
===Fuel Cells===<br />
<br />
Procedure<br />
<br />
# Pipet 75mL of NaPiPES solution into each side of fuel cells<br />
# Inject 1mL of Resazurin solution into each side of chamber<br />
# Using Luer Lock nozzles, connect tubing from top ports to beaker w/ distilled water<br />
# Cut tubing to span distance from manifolds to each fuel cell (1 from nitrogen, 1 from air)<br />
# Attach syringe needles to tubing via Luer Lock nozzles<br />
# Using Luer Lock nozzles, connect tubing from manifolds<br />
# Start gas flow<br />
# Poke needles through bottom ports on fuel cells<br />
<br />
===Measurements===<br />
<br />
Procedure<br />
<br />
# Turn on computer and digital multimeter<br />
# Open LabVIEW program<br />
# Click Run arrow to take resistance readings<br />
# Connect fuel cells to resistor array via alligator clips<br />
# Click "Begin Current Readings" Icon on instrument display<br />
<br />
===Injections/Variables===<br />
<br />
Procedure<br />
<br />
# Once current readings reach equilibrium, inject bacteria into fuels cells using syringes<br />
# Allow bacteria to consume any carbon sources left in their media (approximately 12 hours)<br />
# Once current levels reach stable baseline, inject 1ml lactate solution<br />
# Inject additional variables as desired<br />
<br />
==Clean Up==<br />
<br />
Procedure<br />
<br />
# Drain chambers and soak in 70% ethanol<br />
# Remove carbon felt from anodes and discard (save titanium)<br />
# Scrub all parts in ethanol and distilled water successively<br />
# Use pipe cleaners on ports and tubes<br />
<br />
<br />
<br />
|}<br />
<br><br><br />
<br />
<!--- end body ---><br />
|}</div>Joyyhttp://2008.igem.org/Team:Harvard/Hardware/MFCProcedureTeam:Harvard/Hardware/MFCProcedure2008-10-30T03:52:20Z<p>Joyy: /* Constructing Fuel Cell Components */</p>
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<!--- body here---><br />
{|align="justify" style="background-color:#FFFFFF;text-indent: 15pt;text-align:justify" cellpadding="50" width="90%"<br />
|<br />
=Running an MFC Experiment=<br />
<br />
This page is intended as a comprehensive guide to completing a microbial fuel cell setup and running an experiment from start to finish. <br />
<br />
==Creating a Testing Environment==<br />
<br />
Begin 1-2 weeks prior to experiment<br />
<br />
===Constructing Fuel Cell Components===<br />
<br />
Materials (per fuel cell)<br />
<br />
* 4" Polycarbonate Square Tube, 2" Outer Diameter<br />
* 6" x 6" Polycarbonate Sheet, 1/4" Thick<br />
* 4 Steel Fully Threaded Stud, 1/4"-20 Thread, 6" Length<br />
* 8 Zinc Alloy Wing Flange Nut, 1/4"-20 Screw Size, 1" Wing Spread<br />
* 1" x 1" Nafion® membrane, 0.180mm thick<br />
* 1" x 1" Carbon felt, 0.25" thick<br />
* 1.5" x 1.5" E-TEK ELAT™ GDE (platinum on carbon)<br />
* 2' Titanium Grade 2 Wire .046" Diameter<br />
* Teflon Tape, 1/4" Width<br />
* 5" x 2.5" Silicone Sheet<br />
* Silicone Glue<br />
* Spiral Point Tap 1/4"-28<br />
* 8 Plastic Luer Lock Coupling Nylon, Female to Male Thread, 1/4"-28<br />
* 8 Luer Lock Injection Ports<br />
<br />
Procedure<br />
<br />
# Mill Polycarbonate<br />
#* Cut polycarbonate sheet into 4 equal 3" x 3" pieces<br />
#* Drill four 3/8" holes through each piece, 1 per corner, indented 5mm from both sides<br />
[[Image:endplates.jpg | 600px]]<br />
#* Cut polycarbonate tube into two equal 2" halves<br />
#* Drill four 1/4" holes through each half in configuration shown<br />
[[Image:drilled_tube.jpg | 600px]]<br />
#* Tap each hole with 1/4" -28 spiral tap<br />
<br />
# Glue Chambers (repeat for each half)<br />
#* Center tube on endplate by marking plate with 'X' from corner to corner<br />
#* Squirt 2mm thick line of silicone on edge of tube (edge furthest from holes)<br />
#* Press tube firmly against marked location on endplate<br />
#* Quickly spread excess silicone along edge<br />
#* Let stand 24h to harden<br />
[[Image:glued_half.jpg | 600px]]<br />
<br />
# Construct Gaskets<br />
#* Cut silicone sheet into two equal 2.25" x 2.25" pieces<br />
#* Cut out centered inner squares in each piece, 1.75" x 1.75"<br />
#* Using inner squares, cut two 'O' rings, inner diameter 1/4", outer diameter 1/2"<br />
[[Image:gaskets.jpg | 600px]]<br />
<br />
# Construct Electrodes<br />
#* Cut titanium wire into one 8" piece and one 16" piece<br />
#* Using pliers, shape anode and cathode as shown<br />
'''Anode Frame'''<br><br />
[[Image:Anodeframe.jpg | 600px]]<br />
<br />
#* Spear carbon felt with tip of anode titanium wire and wedge into frame<br />
#* Weave platinum carbon cloth through cathode titanium wire<br />
[[Image:electrodes.jpg | 600px]]<br />
<br />
# Seal Injection Ports<br />
#* Wrap threads of all eight Luer Lock screws with 1' of teflon tape in opposite direction of screwing<br />
#* Screw Luer Locks into all tapped holes in both chambers<br />
[[Image:fin_chamber.jpg | 600px]]<br />
<br />
===Setup of Measurement Device===<br />
<br />
Materials<br />
* Keithley 2700 Digital Multimeter<br />
* Keithley 7700 Multiplexer<br />
* Small Breadboard<br />
* Supply of insulted thin copper wire<br />
* 470 Ohm resistors (one/fuel cell)<br />
<br />
Procedure<br />
# Wire Multiplexer<br />
#* Open multiplexer, note channels<br />
#* Cut two wire 18" wire leads per fuel cell<br />
#* Strip ends, place one wire in each screw terminal, screw tight<br />
#* Tape paired wires (two are attached to each channel) near non-attached ends and label<br />
#* Clamp wire bundles near back of device with provided plastic latch clamps<br />
#* Close Multiplexer and slide into 2700 DMM<br />
<br />
# Create Resistor Array<br />
#* Connect resistors across middle of breadboard (one per fuel cell)<br />
#* Connect leads from multiplexer across resistors (one pair across each resistor)<br />
<br />
<br />
===Controlling the DMM with LabVIEW™===<br />
<br />
# Initialize Multimeter<br />
#* Attach 2700 to COM1 port of desktop computer w/ LabVIEW™<br />
#* Download our LabVIEW™ source code [[Media:MFCs.txt|MFCs.vi]]<br />
#* Open Program in LabVIEW™, adjust block diagram as necessary<br />
<br />
==Experiment Preparation==<br />
<br />
Begin 1 day prior to experiment<br />
<br />
===Assembling Chambers===<br />
<br />
Procedure<br />
# Prepare Electrodes<br />
#* Attach Luer Lock injection ports to all chamber screws<br />
#* Poke tip of electrodes through designated ports from the inside<br />
[[Image:chamber_w_elec.jpg | 600px]]<br />
<br />
# Align Chambers<br />
#* Lay one chamber on a flat surface<br />
#* Place silicone square ring on top edge of tube<br />
#* Place polycarbonate square on silicone<br />
#* Place silicone 'O' ring around central pore<br />
#* Place Nafion membrane on top of 'O' ring<br />
#* Sandwich membrane between second 'O' ring<br />
#* Align second polycarbonate square on top of 'O' ring<br />
#* Center second silicone square ring on polycarbonate square<br />
#* Set second chamber on top of silicone, ensuring ports facing same direction as first chamber<br />
<br />
# Clamp Chambers<br />
#* Move assembly into vice or clamp<br />
#* Insert rods through holes in end plates and screw on wing nuts<br />
#* Tighten evenly<br />
[[Image:fuelcells.jpg | 600px]]<br />
<br />
===Solutions Prep===<br />
<br />
# Chamber media (150ml / fuel cell)<br />
#* 5.844 g/L 100mM NaCl<br />
#* 15.1185 g/L 50mM PIPES (hydrogen) <br />
#* ''7.0 pH''<br />
<br />
# Phosphate buffer (60ml / fuel cell)<br />
#* 2.918 g/L Monosodium phosphate, monohydrate<br />
#* 4.095 g/L Disodium phosphate, anhydrous <br />
#* 5.844 g/L 100mM NaCl<br />
#* ''7.0 pH''<br />
<br />
===Gas Tubing Assembly===<br />
<br />
Materials<br />
* 25' Silicone Soft Rubber Tubing, 3/32" ID, 7/32" OD, 1/16" Wall<br />
* Tank of Compressed Nitrogen<br />
* Gas Regulator<br />
* Lab Supply of Air<br />
* 4 Plastic Luer Lock Coupling Nylon, Male X Barb, for 3/32" Tube<br />
* 4 Plastic Luer Lock Coupling Nylon, Female X Barb, for 3/32" Tube<br />
* Plastic Luer Lock Coupling Nylon, T junctions, for 3/32" Tube<br />
* Syringe needles - 27 gauge<br />
* 2 Aspirator Flasks<br />
* 2 Rubber Stoppers<br />
<br />
Procedure<br />
# Make Flow Regulators<br />
#* Insert nozzle of female Luer Lock into rubber stoppers (poke hole if necessary)<br />
#* Cap aspirator flasks with rubber stoppers<br />
#* Attach tubing from gas sources to each glass nozzle of aspirator flask<br />
[[Image:flow_regs.jpg | 600px]]<br />
<br />
# Make Manifolds<br />
#* Attach T-junction Luer Lock pieces into manifold (1 junction/ fuel cell ; 2 manifods total)<br />
#* Turn last juction such that off is facing end of manifold<br />
#* Attach tubing from stopper of flow regulators to beginning of each mainfold<br />
[[Image:manifolds.jpg | 600px]]<br />
<br />
===Growing Strains===<br />
<br />
Materials<br />
<br />
* 150mL LB / strain<br />
* 1000mL airating flask / strain<br />
* Antibiotics (if desired strain must be selected for)<br />
* Plate or Glycerol Stock with desired strain<br />
<br />
Procedure<br />
<br />
# Fill flasks with LB<br />
# Add correct concentration of selection antibiotic<br />
# Pick single colony from plate, add to flask<br />
# Incubate overnight in shaker at 30C<br />
<br />
==Runtime==<br />
<br />
Begin 2 hours prior to experiment<br />
<br />
===Bacteria===<br />
<br />
Procedure<br />
<br />
# Pipet the cells out of the flask and into 250mL centrifuge containers <br />
# Make sure the containers are close in weight (within 0.5g of each other)<br />
# For a culture more than 250mL split it into two containers <br />
# Set the centrifuge temperature to 22-23C, spin speed to 5000RPM, and time to 15 min<br />
# After first spin, drain each container of the LB, making sure to leave the bacteria pellet intact<br />
# Resuspend pellet in 50mL of potassium buffer<br />
# After bacteria are fully resuspended (no pellets at all), spin down again, 22-23C, 5000RPM, 15min.<br />
# Pour potassium buffer out of container slowly.<br />
# Resuspend pellet in 50mL of potassium buffer<br />
# Spin down again, 22-23C, 5000RPM, 15min.<br />
# Pour potassium buffer out of container slowly. <br />
# Resuspend pellet in 4 mL of sodium pipes<br />
# Check OD (100microliters in 15mL or 1:150 dilution)<br />
# If OD is in linear range, calculate dilution for desired quantity of bacteria in 1mL (typically 10^8 cells /mL of chamber media)<br />
# Repeat dilution if not in linear range (using different ratio)<br />
# Inject 1mL of bacteria into each chamber (see below)<br />
<br />
===Fuel Cells===<br />
<br />
Procedure<br />
<br />
# Pipet 75mL of NaPiPES solution into each side of fuel cells<br />
# Inject 1mL of Resazurin solution into each side of chamber<br />
# Using Luer Lock nozzles, connect tubing from top ports to beaker w/ distilled water<br />
# Cut tubing to span distance from manifolds to each fuel cell (1 from nitrogen, 1 from air)<br />
# Attach syringe needles to tubing via Luer Lock nozzles<br />
# Using Luer Lock nozzles, connect tubing from manifolds<br />
# Start gas flow<br />
# Poke needles through bottom ports on fuel cells<br />
<br />
===Measurements===<br />
<br />
Procedure<br />
<br />
# Turn on computer and digital multimeter<br />
# Open LabVIEW program<br />
# Click Run arrow to take resistance readings<br />
# Connect fuel cells to resistor array via alligator clips<br />
# Click "Begin Current Readings" Icon on instrument display<br />
<br />
===Injections/Variables===<br />
<br />
Procedure<br />
<br />
# Once current readings reach equilibrium, inject bacteria into fuels cells using syringes<br />
# Allow bacteria to consume any carbon sources left in their media (approximately 12 hours)<br />
# Once current levels reach stable baseline, inject 1ml lactate solution<br />
# Inject additional variables as desired<br />
<br />
==Clean Up==<br />
<br />
Procedure<br />
<br />
# Drain chambers and soak in 70% ethanol<br />
# Remove carbon felt from anodes and discard (save titanium)<br />
# Scrub all parts in ethanol and distilled water successively<br />
# Use pipe cleaners on ports and tubes<br />
<br />
<br />
<br />
|}<br />
<br><br><br />
<br />
<!--- end body ---><br />
|}</div>Joyyhttp://2008.igem.org/File:Anodeframe.jpgFile:Anodeframe.jpg2008-10-30T03:51:54Z<p>Joyy: </p>
<hr />
<div></div>Joyyhttp://2008.igem.org/Team:Harvard/Hardware/MFCProcedureTeam:Harvard/Hardware/MFCProcedure2008-10-30T03:51:38Z<p>Joyy: /* Creating a Testing Environment */</p>
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{|align="justify" style="background-color:#FFFFFF;text-indent: 15pt;text-align:justify" cellpadding="50" width="90%"<br />
|<br />
=Running an MFC Experiment=<br />
<br />
This page is intended as a comprehensive guide to completing a microbial fuel cell setup and running an experiment from start to finish. <br />
<br />
==Creating a Testing Environment==<br />
<br />
Begin 1-2 weeks prior to experiment<br />
<br />
===Constructing Fuel Cell Components===<br />
<br />
Materials (per fuel cell)<br />
<br />
* 4" Polycarbonate Square Tube, 2" Outer Diameter<br />
* 6" x 6" Polycarbonate Sheet, 1/4" Thick<br />
* 4 Steel Fully Threaded Stud, 1/4"-20 Thread, 6" Length<br />
* 8 Zinc Alloy Wing Flange Nut, 1/4"-20 Screw Size, 1" Wing Spread<br />
* 1" x 1" Nafion® membrane, 0.180mm thick<br />
* 1" x 1" Carbon felt, 0.25" thick<br />
* 1.5" x 1.5" E-TEK ELAT™ GDE (platinum on carbon)<br />
* 2' Titanium Grade 2 Wire .046" Diameter<br />
* Teflon Tape, 1/4" Width<br />
* 5" x 2.5" Silicone Sheet<br />
* Silicone Glue<br />
* Spiral Point Tap 1/4"-28<br />
* 8 Plastic Luer Lock Coupling Nylon, Female to Male Thread, 1/4"-28<br />
* 8 Luer Lock Injection Ports<br />
<br />
Procedure<br />
<br />
# Mill Polycarbonate<br />
#* Cut polycarbonate sheet into 4 equal 3" x 3" pieces<br />
#* Drill four 3/8" holes through each piece, 1 per corner, indented 5mm from both sides<br />
[[Image:endplates.jpg | 600px]]<br />
#* Cut polycarbonate tube into two equal 2" halves<br />
#* Drill four 1/4" holes through each half in configuration shown<br />
[[Image:drilled_tube.jpg | 600px]]<br />
#* Tap each hole with 1/4" -28 spiral tap<br />
<br />
# Glue Chambers (repeat for each half)<br />
#* Center tube on endplate by marking plate with 'X' from corner to corner<br />
#* Squirt 2mm thick line of silicone on edge of tube (edge furthest from holes)<br />
#* Press tube firmly against marked location on endplate<br />
#* Quickly spread excess silicone along edge<br />
#* Let stand 24h to harden<br />
[[Image:glued_half.jpg | 600px]]<br />
<br />
# Construct Gaskets<br />
#* Cut silicone sheet into two equal 2.25" x 2.25" pieces<br />
#* Cut out centered inner squares in each piece, 1.75" x 1.75"<br />
#* Using inner squares, cut two 'O' rings, inner diameter 1/4", outer diameter 1/2"<br />
[[Image:gaskets.jpg | 600px]]<br />
<br />
# Construct Electrodes<br />
#* Cut titanium wire into one 8" piece and one 16" piece<br />
#* Using pliers, shape anode and cathode as shown<br />
''Anode Frame''<br />
[[Image:Anodeframe.jpg | 600px]]<br />
<br />
#* Spear carbon felt with tip of anode titanium wire and wedge into frame<br />
#* Weave platinum carbon cloth through cathode titanium wire<br />
[[Image:electrodes.jpg | 600px]]<br />
<br />
# Seal Injection Ports<br />
#* Wrap threads of all eight Luer Lock screws with 1' of teflon tape in opposite direction of screwing<br />
#* Screw Luer Locks into all tapped holes in both chambers<br />
[[Image:fin_chamber.jpg | 600px]]<br />
<br />
<br />
===Setup of Measurement Device===<br />
<br />
Materials<br />
* Keithley 2700 Digital Multimeter<br />
* Keithley 7700 Multiplexer<br />
* Small Breadboard<br />
* Supply of insulted thin copper wire<br />
* 470 Ohm resistors (one/fuel cell)<br />
<br />
Procedure<br />
# Wire Multiplexer<br />
#* Open multiplexer, note channels<br />
#* Cut two wire 18" wire leads per fuel cell<br />
#* Strip ends, place one wire in each screw terminal, screw tight<br />
#* Tape paired wires (two are attached to each channel) near non-attached ends and label<br />
#* Clamp wire bundles near back of device with provided plastic latch clamps<br />
#* Close Multiplexer and slide into 2700 DMM<br />
<br />
# Create Resistor Array<br />
#* Connect resistors across middle of breadboard (one per fuel cell)<br />
#* Connect leads from multiplexer across resistors (one pair across each resistor)<br />
<br />
<br />
===Controlling the DMM with LabVIEW™===<br />
<br />
# Initialize Multimeter<br />
#* Attach 2700 to COM1 port of desktop computer w/ LabVIEW™<br />
#* Download our LabVIEW™ source code [[Media:MFCs.txt|MFCs.vi]]<br />
#* Open Program in LabVIEW™, adjust block diagram as necessary<br />
<br />
==Experiment Preparation==<br />
<br />
Begin 1 day prior to experiment<br />
<br />
===Assembling Chambers===<br />
<br />
Procedure<br />
# Prepare Electrodes<br />
#* Attach Luer Lock injection ports to all chamber screws<br />
#* Poke tip of electrodes through designated ports from the inside<br />
[[Image:chamber_w_elec.jpg | 600px]]<br />
<br />
# Align Chambers<br />
#* Lay one chamber on a flat surface<br />
#* Place silicone square ring on top edge of tube<br />
#* Place polycarbonate square on silicone<br />
#* Place silicone 'O' ring around central pore<br />
#* Place Nafion membrane on top of 'O' ring<br />
#* Sandwich membrane between second 'O' ring<br />
#* Align second polycarbonate square on top of 'O' ring<br />
#* Center second silicone square ring on polycarbonate square<br />
#* Set second chamber on top of silicone, ensuring ports facing same direction as first chamber<br />
<br />
# Clamp Chambers<br />
#* Move assembly into vice or clamp<br />
#* Insert rods through holes in end plates and screw on wing nuts<br />
#* Tighten evenly<br />
[[Image:fuelcells.jpg | 600px]]<br />
<br />
===Solutions Prep===<br />
<br />
# Chamber media (150ml / fuel cell)<br />
#* 5.844 g/L 100mM NaCl<br />
#* 15.1185 g/L 50mM PIPES (hydrogen) <br />
#* ''7.0 pH''<br />
<br />
# Phosphate buffer (60ml / fuel cell)<br />
#* 2.918 g/L Monosodium phosphate, monohydrate<br />
#* 4.095 g/L Disodium phosphate, anhydrous <br />
#* 5.844 g/L 100mM NaCl<br />
#* ''7.0 pH''<br />
<br />
===Gas Tubing Assembly===<br />
<br />
Materials<br />
* 25' Silicone Soft Rubber Tubing, 3/32" ID, 7/32" OD, 1/16" Wall<br />
* Tank of Compressed Nitrogen<br />
* Gas Regulator<br />
* Lab Supply of Air<br />
* 4 Plastic Luer Lock Coupling Nylon, Male X Barb, for 3/32" Tube<br />
* 4 Plastic Luer Lock Coupling Nylon, Female X Barb, for 3/32" Tube<br />
* Plastic Luer Lock Coupling Nylon, T junctions, for 3/32" Tube<br />
* Syringe needles - 27 gauge<br />
* 2 Aspirator Flasks<br />
* 2 Rubber Stoppers<br />
<br />
Procedure<br />
# Make Flow Regulators<br />
#* Insert nozzle of female Luer Lock into rubber stoppers (poke hole if necessary)<br />
#* Cap aspirator flasks with rubber stoppers<br />
#* Attach tubing from gas sources to each glass nozzle of aspirator flask<br />
[[Image:flow_regs.jpg | 600px]]<br />
<br />
# Make Manifolds<br />
#* Attach T-junction Luer Lock pieces into manifold (1 junction/ fuel cell ; 2 manifods total)<br />
#* Turn last juction such that off is facing end of manifold<br />
#* Attach tubing from stopper of flow regulators to beginning of each mainfold<br />
[[Image:manifolds.jpg | 600px]]<br />
<br />
===Growing Strains===<br />
<br />
Materials<br />
<br />
* 150mL LB / strain<br />
* 1000mL airating flask / strain<br />
* Antibiotics (if desired strain must be selected for)<br />
* Plate or Glycerol Stock with desired strain<br />
<br />
Procedure<br />
<br />
# Fill flasks with LB<br />
# Add correct concentration of selection antibiotic<br />
# Pick single colony from plate, add to flask<br />
# Incubate overnight in shaker at 30C<br />
<br />
==Runtime==<br />
<br />
Begin 2 hours prior to experiment<br />
<br />
===Bacteria===<br />
<br />
Procedure<br />
<br />
# Pipet the cells out of the flask and into 250mL centrifuge containers <br />
# Make sure the containers are close in weight (within 0.5g of each other)<br />
# For a culture more than 250mL split it into two containers <br />
# Set the centrifuge temperature to 22-23C, spin speed to 5000RPM, and time to 15 min<br />
# After first spin, drain each container of the LB, making sure to leave the bacteria pellet intact<br />
# Resuspend pellet in 50mL of potassium buffer<br />
# After bacteria are fully resuspended (no pellets at all), spin down again, 22-23C, 5000RPM, 15min.<br />
# Pour potassium buffer out of container slowly.<br />
# Resuspend pellet in 50mL of potassium buffer<br />
# Spin down again, 22-23C, 5000RPM, 15min.<br />
# Pour potassium buffer out of container slowly. <br />
# Resuspend pellet in 4 mL of sodium pipes<br />
# Check OD (100microliters in 15mL or 1:150 dilution)<br />
# If OD is in linear range, calculate dilution for desired quantity of bacteria in 1mL (typically 10^8 cells /mL of chamber media)<br />
# Repeat dilution if not in linear range (using different ratio)<br />
# Inject 1mL of bacteria into each chamber (see below)<br />
<br />
===Fuel Cells===<br />
<br />
Procedure<br />
<br />
# Pipet 75mL of NaPiPES solution into each side of fuel cells<br />
# Inject 1mL of Resazurin solution into each side of chamber<br />
# Using Luer Lock nozzles, connect tubing from top ports to beaker w/ distilled water<br />
# Cut tubing to span distance from manifolds to each fuel cell (1 from nitrogen, 1 from air)<br />
# Attach syringe needles to tubing via Luer Lock nozzles<br />
# Using Luer Lock nozzles, connect tubing from manifolds<br />
# Start gas flow<br />
# Poke needles through bottom ports on fuel cells<br />
<br />
===Measurements===<br />
<br />
Procedure<br />
<br />
# Turn on computer and digital multimeter<br />
# Open LabVIEW program<br />
# Click Run arrow to take resistance readings<br />
# Connect fuel cells to resistor array via alligator clips<br />
# Click "Begin Current Readings" Icon on instrument display<br />
<br />
===Injections/Variables===<br />
<br />
Procedure<br />
<br />
# Once current readings reach equilibrium, inject bacteria into fuels cells using syringes<br />
# Allow bacteria to consume any carbon sources left in their media (approximately 12 hours)<br />
# Once current levels reach stable baseline, inject 1ml lactate solution<br />
# Inject additional variables as desired<br />
<br />
==Clean Up==<br />
<br />
Procedure<br />
<br />
# Drain chambers and soak in 70% ethanol<br />
# Remove carbon felt from anodes and discard (save titanium)<br />
# Scrub all parts in ethanol and distilled water successively<br />
# Use pipe cleaners on ports and tubes<br />
<br />
<br />
<br />
|}<br />
<br><br><br />
<br />
<!--- end body ---><br />
|}</div>Joyyhttp://2008.igem.org/File:AnodeFrame.jpgFile:AnodeFrame.jpg2008-10-30T03:50:30Z<p>Joyy: </p>
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<div></div>Joyyhttp://2008.igem.org/File:Gaskets.jpgFile:Gaskets.jpg2008-10-30T03:50:03Z<p>Joyy: </p>
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<div></div>Joyyhttp://2008.igem.org/File:Glued_half.jpgFile:Glued half.jpg2008-10-30T03:49:35Z<p>Joyy: </p>
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<div></div>Joyyhttp://2008.igem.org/File:Drilled_tube.jpgFile:Drilled tube.jpg2008-10-30T03:48:15Z<p>Joyy: </p>
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<div></div>Joyyhttp://2008.igem.org/File:Endplates.jpgFile:Endplates.jpg2008-10-30T03:47:46Z<p>Joyy: </p>
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<div></div>Joyyhttp://2008.igem.org/Team:HarvardTeam:Harvard2008-10-30T03:38:39Z<p>Joyy: </p>
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<div style="position: absolute; left: 100px; top: 880px; height: 400px; width: 100px; padding: 1em;"><br />
<img src="https://static.igem.org/mediawiki/2008/0/06/Thanks_border.jpg"><br />
</div><br />
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<br />
<div style="position: absolute; left: 85px; top: 950px; height: 400px; width: 400px; padding: 1em;"><br />
<center><br />
<font style="line-height:170%"><br />
Alain Viel,<br><br />
Orianna Bretschger,<br />
<br>Daad Saffarini,<br />
<br>Helen White,<br />
<br>Remy Chait,<br />
<br>Natalie Farny,<br />
<br>Christina Agapakis,<br />
<br>Jason Lohmueller,<br />
<br>Kim de Mora,<br />
<br>Colleen Hansel,<br />
<br>Peter Girguis,<br />
<br>Christopher Marx,<br />
<br>George Church,<br />
<br>Jagesh V. Shah,<br />
<br>Pam Silver,<br />
<br>Tamara Brenner,<br />
<br>Harvard BioLabs<br />
</font><br />
</center><br />
</div><br />
<br />
<br />
<br />
<br />
<br />
<div style="position: absolute; left: 580px; top: 300px; width:300px; padding: 1em;"><br />
<a href="https://2008.igem.org/Team:Harvard/Project"><br />
<img src="https://static.igem.org/mediawiki/2008/d/d3/Bactricitynutshell.jpg"></a><br />
</div><br />
<div style="position: absolute; left: 640px; top: 495px; width:180px; padding: .5em;"><br />
<font size=1>Our project sought to combine the detecting capabilities of bacteria with the speed and ubiquity of electricity by creating an inducible system in Shewanella oneidensis MR-1 with an electrical output, allowing for the direct integration of this biosensor with electrical circuits via microbial fuel cells.</font><br />
</div><br />
<br />
<br />
<div style="position: absolute; left: 480px; top: 700px; padding: 1em;"><br />
<a href="https://2008.igem.org/Team:Harvard/Shewie"><br />
<img src="https://static.igem.org/mediawiki/2008/6/65/Mainshewie.gif"></a><br />
</div><br />
<div style="position: absolute; left: 510px; top: 800px; width=200px; padding: 1em;"><br />
<font size=1><br />
Shewanella oneidensis MR-1 <br><br />
(fondly referred to as Shewie)<br><br />
is a metabolically versatile, <br><br />
and genetically tractable, gram-<br><br />
negative facultative anaerobe which under <br><br />
anaerobic conditions reduces a number of electron <br><br />
acceptors. This ability can be harnessed by <br><br />
microbial fuel cells to produce an electric current.<br />
</font><br />
</div><br />
<div style="position: absolute; left: 480px; top: 1000px; padding: 1em;"><br />
<a href ="https://2008.igem.org/Team:Harvard/Hardware"><br />
<img src="https://static.igem.org/mediawiki/2008/d/d8/Fuelcellfun.gif"><br />
</a><br />
</div><br />
<div style="position: absolute; left: 550px; top: 1200px; width:200px; padding: 1em;"><br />
<font size=1><br />
The broad goal of our project was to engineer S. Oneidensis to produce a detectable change in electric current in response to some environmental stimulus. In order to observe such a reaction, our first task was to design an environment capable of housing bacteria and measuring current production. The answer? Microbial fuel cells.<br />
</div><br />
</div><br />
<div style="position: absolute; left: 100px; top: 800px; padding: 1em;"><br />
<img src="https://static.igem.org/mediawiki/2008/d/da/Igemthanks.jpg"><br />
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<font color=#ffffff>ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss </font><br />
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|}</div>Joyyhttp://2008.igem.org/Team:HarvardTeam:Harvard2008-10-30T03:37:58Z<p>Joyy: </p>
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<div><html><br />
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table {<br />
background-color: #c4dbea;<br />
font-color: #cccccc;<br />
color:white;<br />
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background-color: #c4dbea;<br />
color: white;<br />
width: 12em;<br />
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<div style="position: absolute; left: 60px; top: 320px; height: 400px; width: 100px; padding: 1em;"><br />
<img src="https://static.igem.org/mediawiki/igem.org/c/cc/Bactricity.jpg"><br />
</div><br />
<br />
<div style="position: absolute; left: 100px; top: 880px; height: 400px; width: 100px; padding: 1em;"><br />
<img src="https://static.igem.org/mediawiki/2008/0/06/Thanks_border.jpg"><br />
</div><br />
<br />
<br />
<br />
<br />
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<br />
<div style="position: absolute; left: 85px; top: 950px; height: 400px; width: 400px; padding: 1em;"><br />
<center><br />
<font style="line-height:200%"><br />
Alain Viel, <br><br />
Orianna Bretschger, <br />
<br>Daad Saffarini, <br />
<br>Helen White, <br />
<br>Remy Chait, <br />
<br>Natalie Farny, <br />
<br>Christina Agapakis, <br />
<br>Jason Lohmueller, <br />
<br>Kim de Mora, <br />
<br>Colleen Hansel, <br />
<br>Peter Girguis, <br />
<br>Christopher Marx, <br />
<br>George Church, <br />
<br>Jagesh V. Shah, <br />
<br>Pam Silver, <br />
<br>Tamara Brenner, <br />
<br>Harvard BioLabs <br />
</font><br />
</center><br />
</div><br />
<br />
<br />
<br />
<br />
<br />
<div style="position: absolute; left: 580px; top: 300px; width:300px; padding: 1em;"><br />
<a href="https://2008.igem.org/Team:Harvard/Project"><br />
<img src="https://static.igem.org/mediawiki/2008/d/d3/Bactricitynutshell.jpg"></a><br />
</div><br />
<div style="position: absolute; left: 640px; top: 495px; width:180px; padding: .5em;"><br />
<font size=1>Our project sought to combine the detecting capabilities of bacteria with the speed and ubiquity of electricity by creating an inducible system in Shewanella oneidensis MR-1 with an electrical output, allowing for the direct integration of this biosensor with electrical circuits via microbial fuel cells.</font><br />
</div><br />
<br />
<br />
<div style="position: absolute; left: 480px; top: 700px; padding: 1em;"><br />
<a href="https://2008.igem.org/Team:Harvard/Shewie"><br />
<img src="https://static.igem.org/mediawiki/2008/6/65/Mainshewie.gif"></a><br />
</div><br />
<div style="position: absolute; left: 510px; top: 800px; width=200px; padding: 1em;"><br />
<font size=1><br />
Shewanella oneidensis MR-1 <br><br />
(fondly referred to as Shewie)<br><br />
is a metabolically versatile, <br><br />
and genetically tractable, gram-<br><br />
negative facultative anaerobe which under <br><br />
anaerobic conditions reduces a number of electron <br><br />
acceptors. This ability can be harnessed by <br><br />
microbial fuel cells to produce an electric current.<br />
</font><br />
</div><br />
<div style="position: absolute; left: 480px; top: 1000px; padding: 1em;"><br />
<a href ="https://2008.igem.org/Team:Harvard/Hardware"><br />
<img src="https://static.igem.org/mediawiki/2008/d/d8/Fuelcellfun.gif"><br />
</a><br />
</div><br />
<div style="position: absolute; left: 550px; top: 1200px; width:200px; padding: 1em;"><br />
<font size=1><br />
The broad goal of our project was to engineer S. Oneidensis to produce a detectable change in electric current in response to some environmental stimulus. In order to observe such a reaction, our first task was to design an environment capable of housing bacteria and measuring current production. The answer? Microbial fuel cells.<br />
</div><br />
</div><br />
<div style="position: absolute; left: 100px; top: 800px; padding: 1em;"><br />
<img src="https://static.igem.org/mediawiki/2008/d/da/Igemthanks.jpg"><br />
</div><br />
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</html><br />
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|}</div>Joyyhttp://2008.igem.org/Team:HarvardTeam:Harvard2008-10-30T03:36:26Z<p>Joyy: </p>
<hr />
<div><html><br />
<head><br />
<style><br />
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</head><br />
<br />
<div style="position: absolute; left: 60px; top: 320px; height: 400px; width: 100px; padding: 1em;"><br />
<img src="https://static.igem.org/mediawiki/igem.org/c/cc/Bactricity.jpg"><br />
</div><br />
<br />
<div style="position: absolute; left: 100px; top: 880px; height: 400px; width: 100px; padding: 1em;"><br />
<img src="https://static.igem.org/mediawiki/2008/0/06/Thanks_border.jpg"><br />
</div><br />
<br />
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<br />
<br />
<br />
<br />
<div style="position: absolute; left: 85px; top: 950px; height: 400px; width: 400px; padding: 1em; line-spacing:2em"><br />
<center><br />
Alain Viel, <br><br />
Orianna Bretschger, <br />
<br>Daad Saffarini, <br />
<br>Helen White, <br />
<br>Remy Chait, <br />
<br>Natalie Farny, <br />
<br>Christina Agapakis, <br />
<br>Jason Lohmueller, <br />
<br>Kim de Mora, <br />
<br>Colleen Hansel, <br />
<br>Peter Girguis, <br />
<br>Christopher Marx, <br />
<br>George Church, <br />
<br>Jagesh V. Shah, <br />
<br>Pam Silver, <br />
<br>Tamara Brenner, <br />
<br>Harvard BioLabs <br />
</center><br />
</div><br />
<br />
<br />
<br />
<br />
<br />
<div style="position: absolute; left: 580px; top: 300px; width:300px; padding: 1em;"><br />
<a href="https://2008.igem.org/Team:Harvard/Project"><br />
<img src="https://static.igem.org/mediawiki/2008/d/d3/Bactricitynutshell.jpg"></a><br />
</div><br />
<div style="position: absolute; left: 640px; top: 495px; width:180px; padding: .5em;"><br />
<font size=1>Our project sought to combine the detecting capabilities of bacteria with the speed and ubiquity of electricity by creating an inducible system in Shewanella oneidensis MR-1 with an electrical output, allowing for the direct integration of this biosensor with electrical circuits via microbial fuel cells.</font><br />
</div><br />
<br />
<br />
<div style="position: absolute; left: 480px; top: 700px; padding: 1em;"><br />
<a href="https://2008.igem.org/Team:Harvard/Shewie"><br />
<img src="https://static.igem.org/mediawiki/2008/6/65/Mainshewie.gif"></a><br />
</div><br />
<div style="position: absolute; left: 510px; top: 800px; width=200px; padding: 1em;"><br />
<font size=1><br />
Shewanella oneidensis MR-1 <br><br />
(fondly referred to as Shewie)<br><br />
is a metabolically versatile, <br><br />
and genetically tractable, gram-<br><br />
negative facultative anaerobe which under <br><br />
anaerobic conditions reduces a number of electron <br><br />
acceptors. This ability can be harnessed by <br><br />
microbial fuel cells to produce an electric current.<br />
</font><br />
</div><br />
<div style="position: absolute; left: 480px; top: 1000px; padding: 1em;"><br />
<a href ="https://2008.igem.org/Team:Harvard/Hardware"><br />
<img src="https://static.igem.org/mediawiki/2008/d/d8/Fuelcellfun.gif"><br />
</a><br />
</div><br />
<div style="position: absolute; left: 550px; top: 1200px; width:200px; padding: 1em;"><br />
<font size=1><br />
The broad goal of our project was to engineer S. Oneidensis to produce a detectable change in electric current in response to some environmental stimulus. In order to observe such a reaction, our first task was to design an environment capable of housing bacteria and measuring current production. The answer? Microbial fuel cells.<br />
</div><br />
</div><br />
<div style="position: absolute; left: 100px; top: 800px; padding: 1em;"><br />
<img src="https://static.igem.org/mediawiki/2008/d/da/Igemthanks.jpg"><br />
</div><br />
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<font color=#ffffff>ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss </font><br />
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|}</div>Joyyhttp://2008.igem.org/Team:HarvardTeam:Harvard2008-10-30T03:36:10Z<p>Joyy: </p>
<hr />
<div><html><br />
<head><br />
<style><br />
table {<br />
background-color: #c4dbea;<br />
font-color: #cccccc;<br />
color:white;<br />
}<br />
a.menu {<br />
background-color: #c4dbea;<br />
color: white;<br />
width: 12em;<br />
}<br />
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.firstHeading {<br />
color:white;<br />
}<br />
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background-color: #c4dbea;<br />
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p {<br />
color:#333333;<br />
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body {<br />
background-color:#c4dbea;<br />
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.firstHeading {<br />
display:none;<br />
}<br />
</style><br />
</head><br />
<br />
<div style="position: absolute; left: 60px; top: 320px; height: 400px; width: 100px; padding: 1em;"><br />
<img src="https://static.igem.org/mediawiki/igem.org/c/cc/Bactricity.jpg"><br />
</div><br />
<br />
<div style="position: absolute; left: 100px; top: 880px; height: 400px; width: 100px; padding: 1em;"><br />
<img src="https://static.igem.org/mediawiki/2008/0/06/Thanks_border.jpg"><br />
</div><br />
<br />
<br />
<br />
<br />
<br />
<br />
<div style="position: absolute; left: 85px; top: 950px; height: 400px; width: 400px; padding: 3em;"><br />
<center><br />
Alain Viel, <br><br />
Orianna Bretschger, <br />
<br>Daad Saffarini, <br />
<br>Helen White, <br />
<br>Remy Chait, <br />
<br>Natalie Farny, <br />
<br>Christina Agapakis, <br />
<br>Jason Lohmueller, <br />
<br>Kim de Mora, <br />
<br>Colleen Hansel, <br />
<br>Peter Girguis, <br />
<br>Christopher Marx, <br />
<br>George Church, <br />
<br>Jagesh V. Shah, <br />
<br>Pam Silver, <br />
<br>Tamara Brenner, <br />
<br>Harvard BioLabs <br />
</center><br />
</div><br />
<br />
<br />
<br />
<br />
<br />
<div style="position: absolute; left: 580px; top: 300px; width:300px; padding: 1em;"><br />
<a href="https://2008.igem.org/Team:Harvard/Project"><br />
<img src="https://static.igem.org/mediawiki/2008/d/d3/Bactricitynutshell.jpg"></a><br />
</div><br />
<div style="position: absolute; left: 640px; top: 495px; width:180px; padding: .5em;"><br />
<font size=1>Our project sought to combine the detecting capabilities of bacteria with the speed and ubiquity of electricity by creating an inducible system in Shewanella oneidensis MR-1 with an electrical output, allowing for the direct integration of this biosensor with electrical circuits via microbial fuel cells.</font><br />
</div><br />
<br />
<br />
<div style="position: absolute; left: 480px; top: 700px; padding: 1em;"><br />
<a href="https://2008.igem.org/Team:Harvard/Shewie"><br />
<img src="https://static.igem.org/mediawiki/2008/6/65/Mainshewie.gif"></a><br />
</div><br />
<div style="position: absolute; left: 510px; top: 800px; width=200px; padding: 1em;"><br />
<font size=1><br />
Shewanella oneidensis MR-1 <br><br />
(fondly referred to as Shewie)<br><br />
is a metabolically versatile, <br><br />
and genetically tractable, gram-<br><br />
negative facultative anaerobe which under <br><br />
anaerobic conditions reduces a number of electron <br><br />
acceptors. This ability can be harnessed by <br><br />
microbial fuel cells to produce an electric current.<br />
</font><br />
</div><br />
<div style="position: absolute; left: 480px; top: 1000px; padding: 1em;"><br />
<a href ="https://2008.igem.org/Team:Harvard/Hardware"><br />
<img src="https://static.igem.org/mediawiki/2008/d/d8/Fuelcellfun.gif"><br />
</a><br />
</div><br />
<div style="position: absolute; left: 550px; top: 1200px; width:200px; padding: 1em;"><br />
<font size=1><br />
The broad goal of our project was to engineer S. Oneidensis to produce a detectable change in electric current in response to some environmental stimulus. In order to observe such a reaction, our first task was to design an environment capable of housing bacteria and measuring current production. The answer? Microbial fuel cells.<br />
</div><br />
</div><br />
<div style="position: absolute; left: 100px; top: 800px; padding: 1em;"><br />
<img src="https://static.igem.org/mediawiki/2008/d/da/Igemthanks.jpg"><br />
</div><br />
<br />
</html><br />
<br />
<br />
{|<br />
| align="center" style="background:#c4dbea"|<br />
<html><a href = "https://2008.igem.org/Team:Harvard"><img src="https://static.igem.org/mediawiki/2008/b/b9/Harvard_logo.png"></a></html><br />
<br><br />
<br />
{{Template:Main}}<br />
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{| style="color:#1b2c8a;background-color:#FFF;" cellpadding="0" cellspacing="0" border="0" bordercolor="#000" width="100%" align="center"|}<br />
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{|align="justify" style="background-color:#FFFFFF;text-indent: 15pt; font-color:#cccccc; text-align:justify" cellpadding="50" width="90%"<br />
|<br />
<font color=#ffffff>ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss </font><br />
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|}<br />
<br><br><br />
<!--- end body ---><br />
|}</div>Joyyhttp://2008.igem.org/Team:HarvardTeam:Harvard2008-10-30T03:35:50Z<p>Joyy: </p>
<hr />
<div><html><br />
<head><br />
<style><br />
table {<br />
background-color: #c4dbea;<br />
font-color: #cccccc;<br />
color:white;<br />
}<br />
a.menu {<br />
background-color: #c4dbea;<br />
color: white;<br />
width: 12em;<br />
}<br />
<br />
.firstHeading {<br />
color:white;<br />
}<br />
<br />
#bodyContent {<br />
background-color: #c4dbea;<br />
}<br />
<br />
#content {<br />
background-color: #c4dbea;<br />
}<br />
<br />
#footer-box {<br />
background-color: #c4dbea;<br />
}<br />
p {<br />
color:#333333;<br />
}<br />
body {<br />
background-color:#c4dbea;<br />
}<br />
.firstHeading {<br />
display:none;<br />
}<br />
</style><br />
</head><br />
<br />
<div style="position: absolute; left: 60px; top: 320px; height: 400px; width: 100px; padding: 1em;"><br />
<img src="https://static.igem.org/mediawiki/igem.org/c/cc/Bactricity.jpg"><br />
</div><br />
<br />
<div style="position: absolute; left: 100px; top: 880px; height: 400px; width: 100px; padding: 1em;"><br />
<img src="https://static.igem.org/mediawiki/2008/0/06/Thanks_border.jpg"><br />
</div><br />
<br />
<br />
<br />
<br />
<br />
<br />
<div style="position: absolute; left: 95px; top: 950px; height: 400px; width: 400px; padding: 1em;"><br />
<center><br />
Alain Viel, <br><br />
Orianna Bretschger, <br />
<br>Daad Saffarini, <br />
<br>Helen White, <br />
<br>Remy Chait, <br />
<br>Natalie Farny, <br />
<br>Christina Agapakis, <br />
<br>Jason Lohmueller, <br />
<br>Kim de Mora, <br />
<br>Colleen Hansel, <br />
<br>Peter Girguis, <br />
<br>Christopher Marx, <br />
<br>George Church, <br />
<br>Jagesh V. Shah, <br />
<br>Pam Silver, <br />
<br>Tamara Brenner, <br />
<br>Harvard BioLabs <br />
</center><br />
</div><br />
<br />
<br />
<br />
<br />
<br />
<div style="position: absolute; left: 580px; top: 300px; width:300px; padding: 1em;"><br />
<a href="https://2008.igem.org/Team:Harvard/Project"><br />
<img src="https://static.igem.org/mediawiki/2008/d/d3/Bactricitynutshell.jpg"></a><br />
</div><br />
<div style="position: absolute; left: 640px; top: 495px; width:180px; padding: .5em;"><br />
<font size=1>Our project sought to combine the detecting capabilities of bacteria with the speed and ubiquity of electricity by creating an inducible system in Shewanella oneidensis MR-1 with an electrical output, allowing for the direct integration of this biosensor with electrical circuits via microbial fuel cells.</font><br />
</div><br />
<br />
<br />
<div style="position: absolute; left: 480px; top: 700px; padding: 1em;"><br />
<a href="https://2008.igem.org/Team:Harvard/Shewie"><br />
<img src="https://static.igem.org/mediawiki/2008/6/65/Mainshewie.gif"></a><br />
</div><br />
<div style="position: absolute; left: 510px; top: 800px; width=200px; padding: 1em;"><br />
<font size=1><br />
Shewanella oneidensis MR-1 <br><br />
(fondly referred to as Shewie)<br><br />
is a metabolically versatile, <br><br />
and genetically tractable, gram-<br><br />
negative facultative anaerobe which under <br><br />
anaerobic conditions reduces a number of electron <br><br />
acceptors. This ability can be harnessed by <br><br />
microbial fuel cells to produce an electric current.<br />
</font><br />
</div><br />
<div style="position: absolute; left: 480px; top: 1000px; padding: 1em;"><br />
<a href ="https://2008.igem.org/Team:Harvard/Hardware"><br />
<img src="https://static.igem.org/mediawiki/2008/d/d8/Fuelcellfun.gif"><br />
</a><br />
</div><br />
<div style="position: absolute; left: 550px; top: 1200px; width:200px; padding: 1em;"><br />
<font size=1><br />
The broad goal of our project was to engineer S. Oneidensis to produce a detectable change in electric current in response to some environmental stimulus. In order to observe such a reaction, our first task was to design an environment capable of housing bacteria and measuring current production. The answer? Microbial fuel cells.<br />
</div><br />
</div><br />
<div style="position: absolute; left: 100px; top: 800px; padding: 1em;"><br />
<img src="https://static.igem.org/mediawiki/2008/d/da/Igemthanks.jpg"><br />
</div><br />
<br />
</html><br />
<br />
<br />
{|<br />
| align="center" style="background:#c4dbea"|<br />
<html><a href = "https://2008.igem.org/Team:Harvard"><img src="https://static.igem.org/mediawiki/2008/b/b9/Harvard_logo.png"></a></html><br />
<br><br />
<br />
{{Template:Main}}<br />
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{| style="color:#1b2c8a;background-color:#FFF;" cellpadding="0" cellspacing="0" border="0" bordercolor="#000" width="100%" align="center"|}<br />
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|}</div>Joyyhttp://2008.igem.org/Team:HarvardTeam:Harvard2008-10-30T03:35:08Z<p>Joyy: </p>
<hr />
<div><html><br />
<head><br />
<style><br />
table {<br />
background-color: #c4dbea;<br />
font-color: #cccccc;<br />
color:white;<br />
}<br />
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</style><br />
</head><br />
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<div style="position: absolute; left: 60px; top: 320px; height: 400px; width: 100px; padding: 1em;"><br />
<img src="https://static.igem.org/mediawiki/igem.org/c/cc/Bactricity.jpg"><br />
</div><br />
<br />
<div style="position: absolute; left: 100px; top: 880px; height: 400px; width: 100px; padding: 1em;"><br />
<img src="https://static.igem.org/mediawiki/2008/0/06/Thanks_border.jpg"><br />
</div><br />
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<br />
<br />
<br />
<div style="position: absolute; left: 100px; top: 950px; height: 400px; width: 400px; padding: 1em;"><br />
<center><br />
Alain Viel, <br />
Orianna Bretschger, <br />
Daad Saffarini, <br />
Helen White, <br />
Remy Chait, <br />
Natalie Farny, <br />
Christina Agapakis, <br />
Jason Lohmueller, <br />
Kim de Mora, <br />
Colleen Hansel, <br />
Peter Girguis, <br />
Christopher Marx, <br />
George Church, <br />
Jagesh V. Shah, <br />
Pam Silver, <br />
Tamara Brenner, <br />
Harvard BioLabs <br />
</center><br />
</div><br />
<br />
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<br />
<br />
<br />
<div style="position: absolute; left: 580px; top: 300px; width:300px; padding: 1em;"><br />
<a href="https://2008.igem.org/Team:Harvard/Project"><br />
<img src="https://static.igem.org/mediawiki/2008/d/d3/Bactricitynutshell.jpg"></a><br />
</div><br />
<div style="position: absolute; left: 640px; top: 495px; width:180px; padding: .5em;"><br />
<font size=1>Our project sought to combine the detecting capabilities of bacteria with the speed and ubiquity of electricity by creating an inducible system in Shewanella oneidensis MR-1 with an electrical output, allowing for the direct integration of this biosensor with electrical circuits via microbial fuel cells.</font><br />
</div><br />
<br />
<br />
<div style="position: absolute; left: 480px; top: 700px; padding: 1em;"><br />
<a href="https://2008.igem.org/Team:Harvard/Shewie"><br />
<img src="https://static.igem.org/mediawiki/2008/6/65/Mainshewie.gif"></a><br />
</div><br />
<div style="position: absolute; left: 510px; top: 800px; width=200px; padding: 1em;"><br />
<font size=1><br />
Shewanella oneidensis MR-1 <br><br />
(fondly referred to as Shewie)<br><br />
is a metabolically versatile, <br><br />
and genetically tractable, gram-<br><br />
negative facultative anaerobe which under <br><br />
anaerobic conditions reduces a number of electron <br><br />
acceptors. This ability can be harnessed by <br><br />
microbial fuel cells to produce an electric current.<br />
</font><br />
</div><br />
<div style="position: absolute; left: 480px; top: 1000px; padding: 1em;"><br />
<a href ="https://2008.igem.org/Team:Harvard/Hardware"><br />
<img src="https://static.igem.org/mediawiki/2008/d/d8/Fuelcellfun.gif"><br />
</a><br />
</div><br />
<div style="position: absolute; left: 550px; top: 1200px; width:200px; padding: 1em;"><br />
<font size=1><br />
The broad goal of our project was to engineer S. Oneidensis to produce a detectable change in electric current in response to some environmental stimulus. In order to observe such a reaction, our first task was to design an environment capable of housing bacteria and measuring current production. The answer? Microbial fuel cells.<br />
</div><br />
</div><br />
<div style="position: absolute; left: 100px; top: 800px; padding: 1em;"><br />
<img src="https://static.igem.org/mediawiki/2008/d/da/Igemthanks.jpg"><br />
</div><br />
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<font color=#ffffff>ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss </font><br />
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|}</div>Joyyhttp://2008.igem.org/Team:HarvardTeam:Harvard2008-10-30T03:34:42Z<p>Joyy: </p>
<hr />
<div><html><br />
<head><br />
<style><br />
table {<br />
background-color: #c4dbea;<br />
font-color: #cccccc;<br />
color:white;<br />
}<br />
a.menu {<br />
background-color: #c4dbea;<br />
color: white;<br />
width: 12em;<br />
}<br />
<br />
.firstHeading {<br />
color:white;<br />
}<br />
<br />
#bodyContent {<br />
background-color: #c4dbea;<br />
}<br />
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}<br />
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}<br />
p {<br />
color:#333333;<br />
}<br />
body {<br />
background-color:#c4dbea;<br />
}<br />
.firstHeading {<br />
display:none;<br />
}<br />
</style><br />
</head><br />
<br />
<div style="position: absolute; left: 60px; top: 320px; height: 400px; width: 100px; padding: 1em;"><br />
<img src="https://static.igem.org/mediawiki/igem.org/c/cc/Bactricity.jpg"><br />
</div><br />
<br />
<div style="position: absolute; left: 100px; top: 880px; height: 400px; width: 100px; padding: 1em;"><br />
<img src="https://static.igem.org/mediawiki/2008/0/06/Thanks_border.jpg"><br />
</div><br />
<br />
<br />
<br />
<br />
<br />
<br />
<div style="position: absolute; left: 100px; top: 880px; height: 400px; width: 100px; padding: 1em;"><br />
<center><br />
Alain Viel, <br />
Orianna Bretschger, <br />
Daad Saffarini, <br />
Helen White, <br />
Remy Chait, <br />
Natalie Farny, <br />
Christina Agapakis, <br />
Jason Lohmueller, <br />
Kim de Mora, <br />
Colleen Hansel, <br />
Peter Girguis, <br />
Christopher Marx, <br />
George Church, <br />
Jagesh V. Shah, <br />
Pam Silver, <br />
Tamara Brenner, <br />
Harvard BioLabs <br />
</center><br />
</div><br />
<br />
<br />
<br />
<br />
<br />
<div style="position: absolute; left: 580px; top: 300px; width:300px; padding: 1em;"><br />
<a href="https://2008.igem.org/Team:Harvard/Project"><br />
<img src="https://static.igem.org/mediawiki/2008/d/d3/Bactricitynutshell.jpg"></a><br />
</div><br />
<div style="position: absolute; left: 640px; top: 495px; width:180px; padding: .5em;"><br />
<font size=1>Our project sought to combine the detecting capabilities of bacteria with the speed and ubiquity of electricity by creating an inducible system in Shewanella oneidensis MR-1 with an electrical output, allowing for the direct integration of this biosensor with electrical circuits via microbial fuel cells.</font><br />
</div><br />
<br />
<br />
<div style="position: absolute; left: 480px; top: 700px; padding: 1em;"><br />
<a href="https://2008.igem.org/Team:Harvard/Shewie"><br />
<img src="https://static.igem.org/mediawiki/2008/6/65/Mainshewie.gif"></a><br />
</div><br />
<div style="position: absolute; left: 510px; top: 800px; width=200px; padding: 1em;"><br />
<font size=1><br />
Shewanella oneidensis MR-1 <br><br />
(fondly referred to as Shewie)<br><br />
is a metabolically versatile, <br><br />
and genetically tractable, gram-<br><br />
negative facultative anaerobe which under <br><br />
anaerobic conditions reduces a number of electron <br><br />
acceptors. This ability can be harnessed by <br><br />
microbial fuel cells to produce an electric current.<br />
</font><br />
</div><br />
<div style="position: absolute; left: 480px; top: 1000px; padding: 1em;"><br />
<a href ="https://2008.igem.org/Team:Harvard/Hardware"><br />
<img src="https://static.igem.org/mediawiki/2008/d/d8/Fuelcellfun.gif"><br />
</a><br />
</div><br />
<div style="position: absolute; left: 550px; top: 1200px; width:200px; padding: 1em;"><br />
<font size=1><br />
The broad goal of our project was to engineer S. Oneidensis to produce a detectable change in electric current in response to some environmental stimulus. In order to observe such a reaction, our first task was to design an environment capable of housing bacteria and measuring current production. The answer? Microbial fuel cells.<br />
</div><br />
</div><br />
<div style="position: absolute; left: 100px; top: 800px; padding: 1em;"><br />
<img src="https://static.igem.org/mediawiki/2008/d/da/Igemthanks.jpg"><br />
</div><br />
<br />
</html><br />
<br />
<br />
{|<br />
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<html><a href = "https://2008.igem.org/Team:Harvard"><img src="https://static.igem.org/mediawiki/2008/b/b9/Harvard_logo.png"></a></html><br />
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{|align="justify" style="background-color:#FFFFFF;text-indent: 15pt; font-color:#cccccc; text-align:justify" cellpadding="50" width="90%"<br />
|<br />
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|}</div>Joyyhttp://2008.igem.org/Team:HarvardTeam:Harvard2008-10-30T03:30:14Z<p>Joyy: </p>
<hr />
<div><html><br />
<head><br />
<style><br />
table {<br />
background-color: #c4dbea;<br />
font-color: #cccccc;<br />
color:white;<br />
}<br />
a.menu {<br />
background-color: #c4dbea;<br />
color: white;<br />
width: 12em;<br />
}<br />
<br />
.firstHeading {<br />
color:white;<br />
}<br />
<br />
#bodyContent {<br />
background-color: #c4dbea;<br />
}<br />
<br />
#content {<br />
background-color: #c4dbea;<br />
}<br />
<br />
#footer-box {<br />
background-color: #c4dbea;<br />
}<br />
p {<br />
color:#333333;<br />
}<br />
body {<br />
background-color:#c4dbea;<br />
}<br />
.firstHeading {<br />
display:none;<br />
}<br />
</style><br />
</head><br />
<br />
<div style="position: absolute; left: 60px; top: 320px; height: 400px; width: 100px; padding: 1em;"><br />
<img src="https://static.igem.org/mediawiki/igem.org/c/cc/Bactricity.jpg"><br />
</div><br />
<br />
<div style="position: absolute; left: 100px; top: 880px; height: 400px; width: 100px; padding: 1em;"><br />
<img src="https://static.igem.org/mediawiki/2008/0/06/Thanks_border.jpg"><br />
</div><br />
<br />
<br />
<div style="position: absolute; left: 580px; top: 300px; width:300px; padding: 1em;"><br />
<a href="https://2008.igem.org/Team:Harvard/Project"><br />
<img src="https://static.igem.org/mediawiki/2008/d/d3/Bactricitynutshell.jpg"></a><br />
</div><br />
<div style="position: absolute; left: 640px; top: 495px; width:180px; padding: .5em;"><br />
<font size=1>Our project sought to combine the detecting capabilities of bacteria with the speed and ubiquity of electricity by creating an inducible system in Shewanella oneidensis MR-1 with an electrical output, allowing for the direct integration of this biosensor with electrical circuits via microbial fuel cells.</font><br />
</div><br />
<br />
<br />
<div style="position: absolute; left: 480px; top: 700px; padding: 1em;"><br />
<a href="https://2008.igem.org/Team:Harvard/Shewie"><br />
<img src="https://static.igem.org/mediawiki/2008/6/65/Mainshewie.gif"></a><br />
</div><br />
<div style="position: absolute; left: 510px; top: 800px; width=200px; padding: 1em;"><br />
<font size=1><br />
Shewanella oneidensis MR-1 <br><br />
(fondly referred to as Shewie)<br><br />
is a metabolically versatile, <br><br />
and genetically tractable, gram-<br><br />
negative facultative anaerobe which under <br><br />
anaerobic conditions reduces a number of electron <br><br />
acceptors. This ability can be harnessed by <br><br />
microbial fuel cells to produce an electric current.<br />
</font><br />
</div><br />
<div style="position: absolute; left: 480px; top: 1000px; padding: 1em;"><br />
<a href ="https://2008.igem.org/Team:Harvard/Hardware"><br />
<img src="https://static.igem.org/mediawiki/2008/d/d8/Fuelcellfun.gif"><br />
</a><br />
</div><br />
<div style="position: absolute; left: 550px; top: 1200px; width:200px; padding: 1em;"><br />
<font size=1><br />
The broad goal of our project was to engineer S. Oneidensis to produce a detectable change in electric current in response to some environmental stimulus. In order to observe such a reaction, our first task was to design an environment capable of housing bacteria and measuring current production. The answer? Microbial fuel cells.<br />
</div><br />
</div><br />
<div style="position: absolute; left: 100px; top: 800px; padding: 1em;"><br />
<img src="https://static.igem.org/mediawiki/2008/d/da/Igemthanks.jpg"><br />
</div><br />
<br />
</html><br />
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{|<br />
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<html><a href = "https://2008.igem.org/Team:Harvard"><img src="https://static.igem.org/mediawiki/2008/b/b9/Harvard_logo.png"></a></html><br />
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|}</div>Joyyhttp://2008.igem.org/Team:HarvardTeam:Harvard2008-10-30T03:29:54Z<p>Joyy: </p>
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<div style="position: absolute; left: 60px; top: 320px; height: 400px; width: 100px; padding: 1em;"><br />
<img src="https://static.igem.org/mediawiki/igem.org/c/cc/Bactricity.jpg"><br />
</div><br />
<br />
<div style="position: absolute; left: 100px; top: 900px; height: 400px; width: 100px; padding: 1em;"><br />
<img src="https://static.igem.org/mediawiki/2008/0/06/Thanks_border.jpg"><br />
</div><br />
<br />
<br />
<div style="position: absolute; left: 580px; top: 300px; width:300px; padding: 1em;"><br />
<a href="https://2008.igem.org/Team:Harvard/Project"><br />
<img src="https://static.igem.org/mediawiki/2008/d/d3/Bactricitynutshell.jpg"></a><br />
</div><br />
<div style="position: absolute; left: 640px; top: 495px; width:180px; padding: .5em;"><br />
<font size=1>Our project sought to combine the detecting capabilities of bacteria with the speed and ubiquity of electricity by creating an inducible system in Shewanella oneidensis MR-1 with an electrical output, allowing for the direct integration of this biosensor with electrical circuits via microbial fuel cells.</font><br />
</div><br />
<br />
<br />
<div style="position: absolute; left: 480px; top: 680px; padding: 1em;"><br />
<a href="https://2008.igem.org/Team:Harvard/Shewie"><br />
<img src="https://static.igem.org/mediawiki/2008/6/65/Mainshewie.gif"></a><br />
</div><br />
<div style="position: absolute; left: 510px; top: 800px; width=200px; padding: 1em;"><br />
<font size=1><br />
Shewanella oneidensis MR-1 <br><br />
(fondly referred to as Shewie)<br><br />
is a metabolically versatile, <br><br />
and genetically tractable, gram-<br><br />
negative facultative anaerobe which under <br><br />
anaerobic conditions reduces a number of electron <br><br />
acceptors. This ability can be harnessed by <br><br />
microbial fuel cells to produce an electric current.<br />
</font><br />
</div><br />
<div style="position: absolute; left: 480px; top: 1000px; padding: 1em;"><br />
<a href ="https://2008.igem.org/Team:Harvard/Hardware"><br />
<img src="https://static.igem.org/mediawiki/2008/d/d8/Fuelcellfun.gif"><br />
</a><br />
</div><br />
<div style="position: absolute; left: 550px; top: 1200px; width:200px; padding: 1em;"><br />
<font size=1><br />
The broad goal of our project was to engineer S. Oneidensis to produce a detectable change in electric current in response to some environmental stimulus. In order to observe such a reaction, our first task was to design an environment capable of housing bacteria and measuring current production. The answer? Microbial fuel cells.<br />
</div><br />
</div><br />
<div style="position: absolute; left: 100px; top: 800px; padding: 1em;"><br />
<img src="https://static.igem.org/mediawiki/2008/d/da/Igemthanks.jpg"><br />
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|}</div>Joyyhttp://2008.igem.org/Team:HarvardTeam:Harvard2008-10-30T03:29:33Z<p>Joyy: </p>
<hr />
<div><html><br />
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<style><br />
table {<br />
background-color: #c4dbea;<br />
font-color: #cccccc;<br />
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background-color: #c4dbea;<br />
color: white;<br />
width: 12em;<br />
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color:white;<br />
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background-color: #c4dbea;<br />
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p {<br />
color:#333333;<br />
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background-color:#c4dbea;<br />
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display:none;<br />
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</style><br />
</head><br />
<br />
<div style="position: absolute; left: 60px; top: 320px; height: 400px; width: 100px; padding: 1em;"><br />
<img src="https://static.igem.org/mediawiki/igem.org/c/cc/Bactricity.jpg"><br />
</div><br />
<br />
<div style="position: absolute; left: 100px; top: 900px; height: 400px; width: 100px; padding: 1em;"><br />
<img src="https://static.igem.org/mediawiki/2008/0/06/Thanks_border.jpg"><br />
</div><br />
<br />
<br />
<div style="position: absolute; left: 580px; top: 300px; width:300px; padding: 1em;"><br />
<a href="https://2008.igem.org/Team:Harvard/Project"><br />
<img src="https://static.igem.org/mediawiki/2008/d/d3/Bactricitynutshell.jpg"></a><br />
</div><br />
<div style="position: absolute; left: 640px; top: 495px; width:180px; padding: .5em;"><br />
<font size=1>Our project sought to combine the detecting capabilities of bacteria with the speed and ubiquity of electricity by creating an inducible system in Shewanella oneidensis MR-1 with an electrical output, allowing for the direct integration of this biosensor with electrical circuits via microbial fuel cells.</font><br />
</div><br />
<br />
<br />
<div style="position: absolute; left: 480px; top: 700px; padding: 1em;"><br />
<a href="https://2008.igem.org/Team:Harvard/Shewie"><br />
<img src="https://static.igem.org/mediawiki/2008/6/65/Mainshewie.gif"></a><br />
</div><br />
<div style="position: absolute; left: 510px; top: 800px; width=200px; padding: 1em;"><br />
<font size=1><br />
Shewanella oneidensis MR-1 <br><br />
(fondly referred to as Shewie)<br><br />
is a metabolically versatile, <br><br />
and genetically tractable, gram-<br><br />
negative facultative anaerobe which under <br><br />
anaerobic conditions reduces a number of electron <br><br />
acceptors. This ability can be harnessed by <br><br />
microbial fuel cells to produce an electric current.<br />
</font><br />
</div><br />
<div style="position: absolute; left: 480px; top: 1000px; padding: 1em;"><br />
<a href ="https://2008.igem.org/Team:Harvard/Hardware"><br />
<img src="https://static.igem.org/mediawiki/2008/d/d8/Fuelcellfun.gif"><br />
</a><br />
</div><br />
<div style="position: absolute; left: 550px; top: 1200px; width:200px; padding: 1em;"><br />
<font size=1><br />
The broad goal of our project was to engineer S. Oneidensis to produce a detectable change in electric current in response to some environmental stimulus. In order to observe such a reaction, our first task was to design an environment capable of housing bacteria and measuring current production. The answer? Microbial fuel cells.<br />
</div><br />
</div><br />
<div style="position: absolute; left: 100px; top: 800px; padding: 1em;"><br />
<img src="https://static.igem.org/mediawiki/2008/d/da/Igemthanks.jpg"><br />
</div><br />
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{|align="justify" style="background-color:#FFFFFF;text-indent: 15pt; font-color:#cccccc; text-align:justify" cellpadding="50" width="90%"<br />
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<font color=#ffffff>ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss </font><br />
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|}</div>Joyyhttp://2008.igem.org/Team:HarvardTeam:Harvard2008-10-30T03:29:06Z<p>Joyy: </p>
<hr />
<div><html><br />
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<style><br />
table {<br />
background-color: #c4dbea;<br />
font-color: #cccccc;<br />
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background-color: #c4dbea;<br />
color: white;<br />
width: 12em;<br />
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.firstHeading {<br />
color:white;<br />
}<br />
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background-color: #c4dbea;<br />
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background-color: #c4dbea;<br />
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background-color: #c4dbea;<br />
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p {<br />
color:#333333;<br />
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body {<br />
background-color:#c4dbea;<br />
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.firstHeading {<br />
display:none;<br />
}<br />
</style><br />
</head><br />
<br />
<div style="position: absolute; left: 60px; top: 320px; height: 400px; width: 100px; padding: 1em;"><br />
<img src="https://static.igem.org/mediawiki/igem.org/c/cc/Bactricity.jpg"><br />
</div><br />
<br />
<div style="position: absolute; left: 120px; top: 900px; height: 400px; width: 100px; padding: 1em;"><br />
<img src="https://static.igem.org/mediawiki/2008/0/06/Thanks_border.jpg"><br />
</div><br />
<br />
<br />
<div style="position: absolute; left: 580px; top: 300px; width:300px; padding: 1em;"><br />
<a href="https://2008.igem.org/Team:Harvard/Project"><br />
<img src="https://static.igem.org/mediawiki/2008/d/d3/Bactricitynutshell.jpg"></a><br />
</div><br />
<div style="position: absolute; left: 640px; top: 495px; width:180px; padding: .5em;"><br />
<font size=1>Our project sought to combine the detecting capabilities of bacteria with the speed and ubiquity of electricity by creating an inducible system in Shewanella oneidensis MR-1 with an electrical output, allowing for the direct integration of this biosensor with electrical circuits via microbial fuel cells.</font><br />
</div><br />
<br />
<br />
<div style="position: absolute; left: 480px; top: 700px; padding: 1em;"><br />
<a href="https://2008.igem.org/Team:Harvard/Shewie"><br />
<img src="https://static.igem.org/mediawiki/2008/6/65/Mainshewie.gif"></a><br />
</div><br />
<div style="position: absolute; left: 510px; top: 800px; width=200px; padding: 1em;"><br />
<font size=1><br />
Shewanella oneidensis MR-1 <br><br />
(fondly referred to as Shewie)<br><br />
is a metabolically versatile, <br><br />
and genetically tractable, gram-<br><br />
negative facultative anaerobe which under <br><br />
anaerobic conditions reduces a number of electron <br><br />
acceptors. This ability can be harnessed by <br><br />
microbial fuel cells to produce an electric current.<br />
</font><br />
</div><br />
<div style="position: absolute; left: 480px; top: 1000px; padding: 1em;"><br />
<a href ="https://2008.igem.org/Team:Harvard/Hardware"><br />
<img src="https://static.igem.org/mediawiki/2008/d/d8/Fuelcellfun.gif"><br />
</a><br />
</div><br />
<div style="position: absolute; left: 550px; top: 1200px; width:200px; padding: 1em;"><br />
<font size=1><br />
The broad goal of our project was to engineer S. Oneidensis to produce a detectable change in electric current in response to some environmental stimulus. In order to observe such a reaction, our first task was to design an environment capable of housing bacteria and measuring current production. The answer? Microbial fuel cells.<br />
</div><br />
</div><br />
<div style="position: absolute; left: 100px; top: 800px; padding: 1em;"><br />
<img src="https://static.igem.org/mediawiki/2008/d/da/Igemthanks.jpg"><br />
</div><br />
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</html><br />
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{|<br />
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<html><a href = "https://2008.igem.org/Team:Harvard"><img src="https://static.igem.org/mediawiki/2008/b/b9/Harvard_logo.png"></a></html><br />
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{|align="justify" style="background-color:#FFFFFF;text-indent: 15pt; font-color:#cccccc; text-align:justify" cellpadding="50" width="90%"<br />
|<br />
<font color=#ffffff>ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss </font><br />
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|}</div>Joyyhttp://2008.igem.org/Team:HarvardTeam:Harvard2008-10-30T03:28:22Z<p>Joyy: </p>
<hr />
<div><html><br />
<head><br />
<style><br />
table {<br />
background-color: #c4dbea;<br />
font-color: #cccccc;<br />
color:white;<br />
}<br />
a.menu {<br />
background-color: #c4dbea;<br />
color: white;<br />
width: 12em;<br />
}<br />
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.firstHeading {<br />
color:white;<br />
}<br />
<br />
#bodyContent {<br />
background-color: #c4dbea;<br />
}<br />
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#content {<br />
background-color: #c4dbea;<br />
}<br />
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#footer-box {<br />
background-color: #c4dbea;<br />
}<br />
p {<br />
color:#333333;<br />
}<br />
body {<br />
background-color:#c4dbea;<br />
}<br />
.firstHeading {<br />
display:none;<br />
}<br />
</style><br />
</head><br />
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<div style="position: absolute; left: 60px; top: 320px; height: 400px; width: 100px; padding: 1em;"><br />
<img src="https://static.igem.org/mediawiki/igem.org/c/cc/Bactricity.jpg"><br />
</div><br />
<br />
<div style="position: absolute; left: 120px; top: 900px; height: 400px; width: 100px; padding: 1em;"><br />
<img src="https://static.igem.org/mediawiki/2008/0/06/Thanks_border.jpg"><br />
</div><br />
<br />
<br />
<div style="position: absolute; left: 580px; top: 300px; width:300px; padding: 1em;"><br />
<a href="https://2008.igem.org/Team:Harvard/Project"><br />
<img src="https://static.igem.org/mediawiki/2008/d/d3/Bactricitynutshell.jpg"></a><br />
</div><br />
<div style="position: absolute; left: 640px; top: 495px; width:180px; padding: .5em;"><br />
<font size=1>Our project sought to combine the detecting capabilities of bacteria with the speed and ubiquity of electricity by creating an inducible system in Shewanella oneidensis MR-1 with an electrical output, allowing for the direct integration of this biosensor with electrical circuits via microbial fuel cells.</font><br />
</div><br />
<br />
<br />
<div style="position: absolute; left: 500px; top: 650px; padding: 1em;"><br />
<a href="https://2008.igem.org/Team:Harvard/Shewie"><br />
<img src="https://static.igem.org/mediawiki/2008/6/65/Mainshewie.gif"></a><br />
</div><br />
<div style="position: absolute; left: 510px; top: 800px; width=200px; padding: 1em;"><br />
<font size=1><br />
Shewanella oneidensis MR-1 <br><br />
(fondly referred to as Shewie)<br><br />
is a metabolically versatile, <br><br />
and genetically tractable, gram-<br><br />
negative facultative anaerobe which under <br><br />
anaerobic conditions reduces a number of electron <br><br />
acceptors. This ability can be harnessed by <br><br />
microbial fuel cells to produce an electric current.<br />
</font><br />
</div><br />
<div style="position: absolute; left: 480px; top: 1000px; padding: 1em;"><br />
<a href ="https://2008.igem.org/Team:Harvard/Hardware"><br />
<img src="https://static.igem.org/mediawiki/2008/d/d8/Fuelcellfun.gif"><br />
</a><br />
</div><br />
<div style="position: absolute; left: 550px; top: 1200px; width:200px; padding: 1em;"><br />
<font size=1><br />
The broad goal of our project was to engineer S. Oneidensis to produce a detectable change in electric current in response to some environmental stimulus. In order to observe such a reaction, our first task was to design an environment capable of housing bacteria and measuring current production. The answer? Microbial fuel cells.<br />
</div><br />
</div><br />
<div style="position: absolute; left: 100px; top: 800px; padding: 1em;"><br />
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<font color=#ffffff>ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss </font><br />
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|}</div>Joyyhttp://2008.igem.org/Team:HarvardTeam:Harvard2008-10-30T03:27:31Z<p>Joyy: </p>
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<a href="https://2008.igem.org/Team:Harvard/Project"><br />
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<div style="position: absolute; left: 640px; top: 495px; width:180px; padding: .5em;"><br />
<font size=1>Our project sought to combine the detecting capabilities of bacteria with the speed and ubiquity of electricity by creating an inducible system in Shewanella oneidensis MR-1 with an electrical output, allowing for the direct integration of this biosensor with electrical circuits via microbial fuel cells.</font><br />
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<div style="position: absolute; left: 590px; top: 650px; padding: 1em;"><br />
<a href="https://2008.igem.org/Team:Harvard/Shewie"><br />
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<div style="position: absolute; left: 510px; top: 800px; width=200px; padding: 1em;"><br />
<font size=1><br />
Shewanella oneidensis MR-1 <br><br />
(fondly referred to as Shewie)<br><br />
is a metabolically versatile, <br><br />
and genetically tractable, gram-<br><br />
negative facultative anaerobe which under <br><br />
anaerobic conditions reduces a number of electron <br><br />
acceptors. This ability can be harnessed by <br><br />
microbial fuel cells to produce an electric current.<br />
</font><br />
</div><br />
<div style="position: absolute; left: 480px; top: 1000px; padding: 1em;"><br />
<a href ="https://2008.igem.org/Team:Harvard/Hardware"><br />
<img src="https://static.igem.org/mediawiki/2008/d/d8/Fuelcellfun.gif"><br />
</a><br />
</div><br />
<div style="position: absolute; left: 550px; top: 1200px; width:200px; padding: 1em;"><br />
<font size=1><br />
The broad goal of our project was to engineer S. Oneidensis to produce a detectable change in electric current in response to some environmental stimulus. In order to observe such a reaction, our first task was to design an environment capable of housing bacteria and measuring current production. The answer? Microbial fuel cells.<br />
</div><br />
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<a href="https://2008.igem.org/Team:Harvard/Project"><br />
<img src="https://static.igem.org/mediawiki/2008/d/d3/Bactricitynutshell.jpg"></a><br />
</div><br />
<div style="position: absolute; left: 640px; top: 495px; width:180px; padding: .5em;"><br />
<font size=1>Our project sought to combine the detecting capabilities of bacteria with the speed and ubiquity of electricity by creating an inducible system in Shewanella oneidensis MR-1 with an electrical output, allowing for the direct integration of this biosensor with electrical circuits via microbial fuel cells.</font><br />
</div><br />
<div style="position: absolute; left: 590px; top: 650px; padding: 1em;"><br />
<a href="https://2008.igem.org/Team:Harvard/Shewie"><br />
<img src="https://static.igem.org/mediawiki/2008/6/65/Mainshewie.gif"></a><br />
</div><br />
<div style="position: absolute; left: 510px; top: 800px; width=200px; padding: 1em;"><br />
<font size=1><br />
Shewanella oneidensis MR-1 <br><br />
(fondly referred to as Shewie)<br><br />
is a metabolically versatile, <br><br />
and genetically tractable, gram-<br><br />
negative facultative anaerobe which under <br><br />
anaerobic conditions reduces a number of electron <br><br />
acceptors. This ability can be harnessed by <br><br />
microbial fuel cells to produce an electric current.<br />
</font><br />
</div><br />
<div style="position: absolute; left: 480px; top: 1000px; padding: 1em;"><br />
<a href ="https://2008.igem.org/Team:Harvard/Hardware"><br />
<img src="https://static.igem.org/mediawiki/2008/d/d8/Fuelcellfun.gif"><br />
</a><br />
</div><br />
<div style="position: absolute; left: 550px; top: 1200px; width:200px; padding: 1em;"><br />
<font size=1><br />
The broad goal of our project was to engineer S. Oneidensis to produce a detectable change in electric current in response to some environmental stimulus. In order to observe such a reaction, our first task was to design an environment capable of housing bacteria and measuring current production. The answer? Microbial fuel cells.<br />
</div><br />
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<div style="position: absolute; left: 100px; top: 800px; padding: 1em;"><br />
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table {<br />
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background-color: #c4dbea;<br />
color: white;<br />
width: 12em;<br />
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color:white;<br />
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p {<br />
color:#333333;<br />
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background-color:#c4dbea;<br />
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<div style="position: absolute; left: 60px; top: 320px; height: 400px; width: 100px; padding: 1em;"><br />
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</div><br />
<div style="position: absolute; left: 580px; top: 300px; width:300px; padding: 1em;"><br />
<a href="https://2008.igem.org/Team:Harvard/Project"><br />
<img src="https://static.igem.org/mediawiki/2008/d/d3/Bactricitynutshell.jpg"></a><br />
</div><br />
<div style="position: absolute; left: 640px; top: 495px; width:180px; padding: .5em;"><br />
<font size=1>Our project sought to combine the detecting capabilities of bacteria with the speed and ubiquity of electricity by creating an inducible system in Shewanella oneidensis MR-1 with an electrical output, allowing for the direct integration of this biosensor with electrical circuits via microbial fuel cells.</font><br />
</div><br />
<div style="position: absolute; left: 490px; top: 700px; padding: 1em;"><br />
<a href="https://2008.igem.org/Team:Harvard/Shewie"><br />
<img src="https://static.igem.org/mediawiki/2008/6/65/Mainshewie.gif"></a><br />
</div><br />
<div style="position: absolute; left: 510px; top: 800px; width=200px; padding: 1em;"><br />
<font size=1><br />
Shewanella oneidensis MR-1 <br><br />
(fondly referred to as Shewie)<br><br />
is a metabolically versatile, <br><br />
and genetically tractable, gram-<br><br />
negative facultative anaerobe which under <br><br />
anaerobic conditions reduces a number of electron <br><br />
acceptors. This ability can be harnessed by <br><br />
microbial fuel cells to produce an electric current.<br />
</font><br />
</div><br />
<div style="position: absolute; left: 480px; top: 1000px; padding: 1em;"><br />
<a href ="https://2008.igem.org/Team:Harvard/Hardware"><br />
<img src="https://static.igem.org/mediawiki/2008/d/d8/Fuelcellfun.gif"><br />
</a><br />
</div><br />
<div style="position: absolute; left: 550px; top: 1200px; width:200px; padding: 1em;"><br />
<font size=1><br />
The broad goal of our project was to engineer S. Oneidensis to produce a detectable change in electric current in response to some environmental stimulus. In order to observe such a reaction, our first task was to design an environment capable of housing bacteria and measuring current production. The answer? Microbial fuel cells.<br />
</div><br />
</div><br />
<div style="position: absolute; left: 100px; top: 800px; padding: 1em;"><br />
<img src="https://static.igem.org/mediawiki/2008/d/da/Igemthanks.jpg"><br />
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<font color=#ffffff>ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss </font><br />
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color: white;<br />
width: 12em;<br />
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.firstHeading {<br />
color:white;<br />
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background-color: #c4dbea;<br />
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p {<br />
color:#333333;<br />
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<img src="https://static.igem.org/mediawiki/igem.org/c/cc/Bactricity.jpg"><br />
</div><br />
<br />
<div style="position: absolute; left: 60px; top: 920px; height: 400px; width: 100px; padding: 1em;"><br />
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<div style="position: absolute; left: 580px; top: 300px; width:300px; padding: 1em;"><br />
<a href="https://2008.igem.org/Team:Harvard/Project"><br />
<img src="https://static.igem.org/mediawiki/2008/d/d3/Bactricitynutshell.jpg"></a><br />
</div><br />
<div style="position: absolute; left: 640px; top: 495px; width:180px; padding: .5em;"><br />
<font size=1>Our project sought to combine the detecting capabilities of bacteria with the speed and ubiquity of electricity by creating an inducible system in Shewanella oneidensis MR-1 with an electrical output, allowing for the direct integration of this biosensor with electrical circuits via microbial fuel cells.</font><br />
</div><br />
<div style="position: absolute; left: 590px; top: 650px; padding: 1em;"><br />
<a href="https://2008.igem.org/Team:Harvard/Shewie"><br />
<img src="https://static.igem.org/mediawiki/2008/6/65/Mainshewie.gif"></a><br />
</div><br />
<div style="position: absolute; left: 510px; top: 800px; width=200px; padding: 1em;"><br />
<font size=1><br />
Shewanella oneidensis MR-1 <br><br />
(fondly referred to as Shewie)<br><br />
is a metabolically versatile, <br><br />
and genetically tractable, gram-<br><br />
negative facultative anaerobe which under <br><br />
anaerobic conditions reduces a number of electron <br><br />
acceptors. This ability can be harnessed by <br><br />
microbial fuel cells to produce an electric current.<br />
</font><br />
</div><br />
<div style="position: absolute; left: 480px; top: 1000px; padding: 1em;"><br />
<a href ="https://2008.igem.org/Team:Harvard/Hardware"><br />
<img src="https://static.igem.org/mediawiki/2008/d/d8/Fuelcellfun.gif"><br />
</a><br />
</div><br />
<div style="position: absolute; left: 550px; top: 1200px; width:200px; padding: 1em;"><br />
<font size=1><br />
The broad goal of our project was to engineer S. Oneidensis to produce a detectable change in electric current in response to some environmental stimulus. In order to observe such a reaction, our first task was to design an environment capable of housing bacteria and measuring current production. The answer? Microbial fuel cells.<br />
</div><br />
</div><br />
<div style="position: absolute; left: 100px; top: 800px; padding: 1em;"><br />
<img src="https://static.igem.org/mediawiki/2008/d/da/Igemthanks.jpg"><br />
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|}</div>Joyyhttp://2008.igem.org/Team:HarvardTeam:Harvard2008-10-30T03:21:09Z<p>Joyy: </p>
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<img src="https://static.igem.org/mediawiki/igem.org/c/cc/Bactricity.jpg"><br />
</div><br />
<br />
<div style="position: absolute; left: 60px; top: 920px; height: 400px; width: 100px; padding: 1em;"><br />
<img src="https://static.igem.org/mediawiki/2008/0/06/Thanks_border.jpg"><br />
</div><br />
<br />
<br />
<div style="position: absolute; left: 580px; top: 300px; width:300px; padding: 1em;"><br />
<a href="https://2008.igem.org/Team:Harvard/Project"><br />
<img src="https://static.igem.org/mediawiki/2008/d/d3/Bactricitynutshell.jpg"></a><br />
</div><br />
<div style="position: absolute; left: 640px; top: 495px; width:180px; padding: .5em;"><br />
<font size=1>Our project sought to combine the detecting capabilities of bacteria with the speed and ubiquity of electricity by creating an inducible system in Shewanella oneidensis MR-1 with an electrical output, allowing for the direct integration of this biosensor with electrical circuits via microbial fuel cells.</font><br />
</div><br />
<div style="position: absolute; left: 490px; top: 700px; padding: 1em;"><br />
<a href="https://2008.igem.org/Team:Harvard/Shewie"><br />
<img src="https://static.igem.org/mediawiki/2008/6/65/Mainshewie.gif"></a><br />
</div><br />
<div style="position: absolute; left: 510px; top: 800px; width=200px; padding: 1em;"><br />
<font size=1><br />
Shewanella oneidensis MR-1 <br><br />
(fondly referred to as Shewie)<br><br />
is a metabolically versatile, <br><br />
and genetically tractable, gram-<br><br />
negative facultative anaerobe which under <br><br />
anaerobic conditions reduces a number of electron <br><br />
acceptors. This ability can be harnessed by <br><br />
microbial fuel cells to produce an electric current.<br />
</font><br />
</div><br />
<div style="position: absolute; left: 480px; top: 1000px; padding: 1em;"><br />
<a href ="https://2008.igem.org/Team:Harvard/Hardware"><br />
<img src="https://static.igem.org/mediawiki/2008/d/d8/Fuelcellfun.gif"><br />
</a><br />
</div><br />
<div style="position: absolute; left: 550px; top: 1200px; width:200px; padding: 1em;"><br />
<font size=1><br />
The broad goal of our project was to engineer S. Oneidensis to produce a detectable change in electric current in response to some environmental stimulus. In order to observe such a reaction, our first task was to design an environment capable of housing bacteria and measuring current production. The answer? Microbial fuel cells.<br />
</div><br />
</div><br />
<div style="position: absolute; left: 100px; top: 800px; padding: 1em;"><br />
<img src="https://static.igem.org/mediawiki/2008/d/da/Igemthanks.jpg"><br />
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<font color=#ffffff>ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss </font><br />
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|}</div>Joyyhttp://2008.igem.org/File:Thanks_border.jpgFile:Thanks border.jpg2008-10-30T03:20:27Z<p>Joyy: </p>
<hr />
<div></div>Joyyhttp://2008.igem.org/Team:HarvardTeam:Harvard2008-10-30T03:16:28Z<p>Joyy: </p>
<hr />
<div><html><br />
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<style><br />
table {<br />
background-color: #c4dbea;<br />
font-color: #cccccc;<br />
color:white;<br />
}<br />
a.menu {<br />
background-color: #c4dbea;<br />
color: white;<br />
width: 12em;<br />
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color:white;<br />
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display:none;<br />
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</head><br />
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<div style="position: absolute; left: 60px; top: 320px; height: 400px; width: 100px; padding: 1em;"><br />
<img src="https://static.igem.org/mediawiki/igem.org/c/cc/Bactricity.jpg"><br />
</div><br />
<div style="position: absolute; left: 580px; top: 300px; width:300px; padding: 1em;"><br />
<a href="https://2008.igem.org/Team:Harvard/Project"><br />
<img src="https://static.igem.org/mediawiki/2008/d/d3/Bactricitynutshell.jpg"></a><br />
</div><br />
<div style="position: absolute; left: 640px; top: 495px; width:180px; padding: .5em;"><br />
<font size=1>Our project sought to combine the detecting capabilities of bacteria with the speed and ubiquity of electricity by creating an inducible system in Shewanella oneidensis MR-1 with an electrical output, allowing for the direct integration of this biosensor with electrical circuits via microbial fuel cells.</font><br />
</div><br />
<div style="position: absolute; left: 490px; top: 700px; padding: 1em;"><br />
<a href="https://2008.igem.org/Team:Harvard/Shewie"><br />
<img src="https://static.igem.org/mediawiki/2008/6/65/Mainshewie.gif"></a><br />
</div><br />
<div style="position: absolute; left: 510px; top: 800px; width=200px; padding: 1em;"><br />
<font size=1><br />
Shewanella oneidensis MR-1 <br><br />
(fondly referred to as Shewie)<br><br />
is a metabolically versatile, <br><br />
and genetically tractable, gram-<br><br />
negative facultative anaerobe which under <br><br />
anaerobic conditions reduces a number of electron <br><br />
acceptors. This ability can be harnessed by <br><br />
microbial fuel cells to produce an electric current.<br />
</font><br />
</div><br />
<div style="position: absolute; left: 480px; top: 1000px; padding: 1em;"><br />
<a href ="https://2008.igem.org/Team:Harvard/Hardware"><br />
<img src="https://static.igem.org/mediawiki/2008/d/d8/Fuelcellfun.gif"><br />
</a><br />
</div><br />
<div style="position: absolute; left: 550px; top: 1200px; width:200px; padding: 1em;"><br />
<font size=1><br />
The broad goal of our project was to engineer S. Oneidensis to produce a detectable change in electric current in response to some environmental stimulus. In order to observe such a reaction, our first task was to design an environment capable of housing bacteria and measuring current production. The answer? Microbial fuel cells.<br />
</div><br />
</div><br />
<div style="position: absolute; left: 100px; top: 800px; padding: 1em;"><br />
<img src="https://static.igem.org/mediawiki/2008/d/da/Igemthanks.jpg"><br />
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<font color=#ffffff>ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss </font><br />
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|}</div>Joyyhttp://2008.igem.org/Team:HarvardTeam:Harvard2008-10-30T03:16:19Z<p>Joyy: </p>
<hr />
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<style><br />
table {<br />
background-color: #c4dbea;<br />
font-color: #cccccc;<br />
color:white;<br />
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background-color: #c4dbea;<br />
color: white;<br />
width: 12em;<br />
}<br />
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color:white;<br />
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p {<br />
color:#333333;<br />
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background-color:#c4dbea;<br />
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display:none;<br />
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</style><br />
</head><br />
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<div style="position: absolute; left: 60px; top: 320px; height: 400px; width: 100px; padding: 1em;"><br />
<img src="https://static.igem.org/mediawiki/igem.org/c/cc/Bactricity.jpg"><br />
</div><br />
<div style="position: absolute; left: 580px; top: 300px; width:300px; padding: 1em;"><br />
<a href="https://2008.igem.org/Team:Harvard/Project"><br />
<img src="https://static.igem.org/mediawiki/2008/d/d3/Bactricitynutshell.jpg"></a><br />
</div><br />
<div style="position: absolute; left: 640px; top: 495px; width:180px; padding: .5em;"><br />
<font size=1>Our project sought to combine the detecting capabilities of bacteria with the speed and ubiquity of electricity by creating an inducible system in Shewanella oneidensis MR-1 with an electrical output, allowing for the direct integration of this biosensor with electrical circuits via microbial fuel cells.</font><br />
</div><br />
<div style="position: absolute; left: 490px; top: 700px; padding: 1em;"><br />
<a href="https://2008.igem.org/Team:Harvard/Shewie"><br />
<img src="https://static.igem.org/mediawiki/2008/6/65/Mainshewie.gif"></a><br />
</div><br />
<div style="position: absolute; left: 510px; top: 800px; width=200px; padding: 1em;"><br />
<font size=1><br />
Shewanella oneidensis MR-1 <br><br />
(fondly referred to as Shewie)<br><br />
is a metabolically versatile, <br><br />
and genetically tractable, gram-<br><br />
negative facultative anaerobe which under <br><br />
anaerobic conditions reduces a number of electron <br><br />
acceptors. This ability can be harnessed by <br><br />
microbial fuel cells to produce an electric current.<br />
</font><br />
</div><br />
<div style="position: absolute; left: 480px; top: 1000px; padding: 1em;"><br />
<a href ="https://2008.igem.org/Team:Harvard/Hardware"><br />
<img src="https://static.igem.org/mediawiki/2008/d/d8/Fuelcellfun.gif"><br />
</a><br />
</div><br />
<div style="position: absolute; left: 550px; top: 1200px; width:100px; padding: 1em;"><br />
<font size=1><br />
The broad goal of our project was to engineer S. Oneidensis to produce a detectable change in electric current in response to some environmental stimulus. In order to observe such a reaction, our first task was to design an environment capable of housing bacteria and measuring current production. The answer? Microbial fuel cells.<br />
</div><br />
</div><br />
<div style="position: absolute; left: 100px; top: 800px; padding: 1em;"><br />
<img src="https://static.igem.org/mediawiki/2008/d/da/Igemthanks.jpg"><br />
</div><br />
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|}</div>Joyyhttp://2008.igem.org/Team:HarvardTeam:Harvard2008-10-30T03:16:01Z<p>Joyy: </p>
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<font size=1>Our project sought to combine the detecting capabilities of bacteria with the speed and ubiquity of electricity by creating an inducible system in Shewanella oneidensis MR-1 with an electrical output, allowing for the direct integration of this biosensor with electrical circuits via microbial fuel cells.</font><br />
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Shewanella oneidensis MR-1 <br><br />
(fondly referred to as Shewie)<br><br />
is a metabolically versatile, <br><br />
and genetically tractable, gram-<br><br />
negative facultative anaerobe which under <br><br />
anaerobic conditions reduces a number of electron <br><br />
acceptors. This ability can be harnessed by <br><br />
microbial fuel cells to produce an electric current.<br />
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The broad goal of our project was to engineer S. Oneidensis to produce a detectable change in electric current in response to some environmental stimulus. In order to observe such a reaction, our first task was to design an environment capable of housing bacteria and measuring current production. The answer? Microbial fuel cells.<br />
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<font color=#ffffff>ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss ssssss </font><br />
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