Team:Lethbridge CCS/Notebook
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2 Sept 2008
(Peter, Paul, Elizabeth, Glenda, Nathan)
- Resuspended our primers (psB1A7 forward and reverse; TetR-GFP forward and reverse) to 100 μM.
- Made 10-fold dilutions of each primer
- Set up and ran PCR on psB1A7
- varied concentration of vector (1 x, 1/10 x, 1/100 x) and annealing temperatures (58°C, 62°C, 66°C), giving nine variations;
- used materials and amounts as per Phusion polymerase kit
- Cycling temperatures and times for PCR Initial denaturation 98°C 30 sec ___ Denaturation 98°C 10 sec | Annealing 58/62/66 °C 30 sec |--- 30 cycles Extension 72°C 1 min ___| Final extension 72°C 10 min Hold 4°C hold
3 Sept 2008
(Peter, Paul, Elizabeth, Marc, Glenda, Nathan)
- PCR of pSB1A7 from yesterday worked well; approximate size was expected to be ~2500 bp, which was verified (see below)
- conditions
6 Sept 2008
- Agarose gel purification of pSB1A7 from [date] (Peter, Paul, Marc, Glenda, Nathan)
- conditions