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Team:University of Ottawa/24 July 2008
From 2008.igem.org
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::<li> Dan and I decided that the initial extraction of PTP2 after PCR yielded a product that was too concentrated to be properly extracted - so I am redoing the PCR amplification with elongation time raised to 1 min 15 sec and annealing temp raised to 58 C. | ::<li> Dan and I decided that the initial extraction of PTP2 after PCR yielded a product that was too concentrated to be properly extracted - so I am redoing the PCR amplification with elongation time raised to 1 min 15 sec and annealing temp raised to 58 C. | ||
::<li> I will also spread the PTP2 amplification product over many different lanes on the gel so that it can be properly extracted. | ::<li> I will also spread the PTP2 amplification product over many different lanes on the gel so that it can be properly extracted. | ||
| + | :'''Digestion''' | ||
| + | ::<li> pSSA42 was redigested with BamHI and xhoI. | ||
Revision as of 18:42, 7 August 2008
Today in the lab
Chris
- Double-Check of AtCRE
- Digested samples showing desired bands with EcoRI for one hour.
- Ran against a water control and DQ2325601 digested with EcoRI for one and a half hours on a 1% gel at 80V
- One of two samples still showed desired bands following double-check. This sample was streaked onto a master plate, which was incubated at 37 C overnight, and glycerol stocked in the -80 C fridge for further use.
Matt
- Ligation Results
- A gel was run after two hours for ligation of PTP2/pSSA42 - unsuccessful.
- A gel was run with overnight ligation - vector and H2O controls were clean and ligation product band is slightly visible in 3:1 ratio. However there is auto ligation occuring with the vector.
- PCR
- Dan and I decided that the initial extraction of PTP2 after PCR yielded a product that was too concentrated to be properly extracted - so I am redoing the PCR amplification with elongation time raised to 1 min 15 sec and annealing temp raised to 58 C.
- I will also spread the PTP2 amplification product over many different lanes on the gel so that it can be properly extracted.
- Digestion
- pSSA42 was redigested with BamHI and xhoI.
