"
Team:Lethbridge CCS/Notebook
From 2008.igem.org
(Difference between revisions)
| Line 22: | Line 22: | ||
| - | === | + | === 3 Sept 2008 === |
| - | (Peter, Paul, Marc, Glenda, Nathan) | + | (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) | * PCR of pSB1A7 from yesterday worked well; approximate size was expected to be ~2500 bp, which was verified (see below) | ||
| - | + | ** [[Image:PCRpSB1A7]] | |
- conditions | - conditions | ||
Revision as of 22:16, 6 September 2008
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
