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Team:University of Ottawa/29 July 2008
From 2008.igem.org
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| - | + | ='''Today in the lab'''= | |
| - | + | __TOC__ | |
| - | '''Chris''' | + | =='''Chris'''== |
:'''PCR Amplification of PTP2''' | :'''PCR Amplification of PTP2''' | ||
::<li> Began running experimentation alongside Matt to increase chances of success | ::<li> Began running experimentation alongside Matt to increase chances of success | ||
| Line 86: | Line 86: | ||
:'''PCR Amplification of PTP2''' | :'''PCR Amplification of PTP2''' | ||
::<li> Ran PCR with same constraints as previously. Let run overnight. | ::<li> Ran PCR with same constraints as previously. Let run overnight. | ||
| + | |||
| + | =='''Tammy and Dan'''== | ||
| + | :'''0.8% Agarose Gel Electrophoresis of T123 PCR Products''' | ||
| + | ::<li> Each PCR reaction tube (50 μL) was divided into 2 and ran on 2 separate wells to decrease DNA load and to ensure identification of appropriate band size | ||
| + | |||
| + | :'''Agarose Gel Extraction''' | ||
| + | ::<li> 24 bands were cut from the gel using minimal UV exposure | ||
| + | ::<li> 6 gel bands were pooled into 1 column for a total of 4 aliquots of purified T123 DNA | ||
| + | |||
| + | =='''Matt'''== | ||
| + | :'''Inoculation''' | ||
| + | ::<li> Inoculation of pSSA42 turned out nicely, control was clean - performed a miniprep from inoculation. | ||
| + | :'''Transformation''' | ||
| + | ::<li> Plates of transformation in competent cells did not yield any colonies - control was clean. | ||
| + | :'''Digestion''' | ||
| + | ::<li> Digestion of pSSA42 was performed with BamHI + xhoI. | ||
Latest revision as of 18:52, 7 August 2008
Today in the lab
Contents |
Chris
- PCR Amplification of PTP2
- Began running experimentation alongside Matt to increase chances of success
- Ran 5 samples, including two water controls as per Cory's request.
- Master mix: 50 uL Buffer, 5 uL dNTP, 12.5 uL F60 and F61, 2.5 uL DNA (at 25 ng/uL), 142.5 uL water.
- Digestion of PSSA42
- Ran five samples and one water control
- Per tube: 3 uL water, 3 uL Buffer 3, 3 uL BSA, 1.5 uL XhoI and BamHI, 16 uL DNA template.
- Incubated at 37 C for one hour
- PCR Cleanup
- Used PCR cleanup kit to purify PTP2
- Measured absorbance of resulting DNA samples. The concentrations were found to be very low; PCR did not work. It was later determined that DNA template was not added, by accident.
- PCR Amplification of PTP2
- Ran PCR with same constraints as previously. Let run overnight.
Tammy and Dan
- 0.8% Agarose Gel Electrophoresis of T123 PCR Products
- Each PCR reaction tube (50 μL) was divided into 2 and ran on 2 separate wells to decrease DNA load and to ensure identification of appropriate band size
- Agarose Gel Extraction
- 24 bands were cut from the gel using minimal UV exposure
- 6 gel bands were pooled into 1 column for a total of 4 aliquots of purified T123 DNA
Matt
- Inoculation
- Inoculation of pSSA42 turned out nicely, control was clean - performed a miniprep from inoculation.
- Transformation
- Plates of transformation in competent cells did not yield any colonies - control was clean.
- Digestion
- Digestion of pSSA42 was performed with BamHI + xhoI.
