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Team:University of Ottawa/29 July 2008
From 2008.igem.org
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.
