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Team:Harvard/Dailybook/Week3/Widgetry
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* Constructed two new MFCs | * Constructed two new MFCs | ||
* grew mtrA, mtrB, wt, and E. coli in LB+lactate liquid cultures | * grew mtrA, mtrB, wt, and E. coli in LB+lactate liquid cultures | ||
| - | * Ran negative control with E. coli in | + | * Ran negative control with E. coli in NaPIPES media |
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| + | |||
| + | [[Image:EColi_NaPIPES.JPG|600px]] | ||
=Wednesday: June 9, 2008= | =Wednesday: June 9, 2008= | ||
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[[Image:Mfc_overnight3.png|600px]] | [[Image:Mfc_overnight3.png|600px]] | ||
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| + | '''Conclusion''' | ||
| + | * Day 1, current production of wt and mtrA Shewie match that in literature | ||
| + | ** When wt peaks at around 275 microamps, mtrA Shewie is producing 25-50 microamps | ||
Latest revision as of 02:10, 30 October 2008
Contents |
Goals for this week
- Make Orianna's growth media
- construct more chambers to run parallel tests
- set up Keithley for multiple tests
- grow mtrA, mtrB,wt, E. coli in liquid cultures
- negative control E. coli in LB/lactate
- standardize tests (OD, bubbling, etc)
Monday: June 7, 2008
- E. Coli Negative Control in LB
Tuesday: June 8, 2008
- Constructed two new MFCs
- grew mtrA, mtrB, wt, and E. coli in LB+lactate liquid cultures
- Ran negative control with E. coli in NaPIPES media
Wednesday: June 9, 2008
- Made Orianna's growth media
- First followed recipe exactly: pH = 3.4
- Added NaOH to neutralize media
- Missing amino acids
- First followed recipe exactly: pH = 3.4
- Grew wt Shewie, mtrA, mtrB, and mtrC/omcA in Orianna's growth media
- overnight cultures
- RESULTS:
- OD readings:
- wt: -0.001
- mtrA: 0.001
- mtrB: 0.003
- mtrC/omcA: 0.002
- CONCLUSION:
- Orianna's growth media isn't working. Go see Colleen for possible sources of error.
- OD readings from liquid cultures grown in LB + lactate growth media
- wt: 2.489
- mtrB: 1.711
- Ran MFC overnight with mtrB and wt Shewie strains.
- added strains first without lactate
- waited until chambers became anaerobic (color-change) to add lactate
- wt cell chamber denser than mtrB cell chamber
- N2 stopped in wt chamber. turned off N2 in mtrB chamber to mirror this change
- Experiment continued
- replenished lactate in wt chamber
- fluctuations may have resulted from air/N2 flow
Thursday: June 10, 2008
- Continued running experiment from Wednesday
- Purchased pipe cleaners, resistors
- Made OGM with potassium PIPES
Friday: June 11, 2008
Lab
- set up three chamber system
- tested wt, mtrA, negative control:
- resistances:
- wt:
- mtrA:
- none:
- resistances:
465 s: injected Shewie
3500 s: injected Lactate & stopped N2
- Nitrogen stopped bubbling somewhere between 70,000 & 80,000
Conclusion
- Day 1, current production of wt and mtrA Shewie match that in literature
- When wt peaks at around 275 microamps, mtrA Shewie is producing 25-50 microamps
