August 18th, 2010
1. Make colony PCR to verify 1) L1: p30-MinBP + GFP E0240 (6 colonies); 2) L2: p30-MinBP + GFP BBa_K145015 (74 min) (10 colonies); 3) L4: Backbone pSB3K3 + Blue Promoter BBa_K238013 + GFP BBa_K145015 (74 min) (3 colonies).
- Tubes are marked as follows:
- -2.1-2.10 -> L2: p30-MinBP + GFP BBa_K145015 (74 min) (10 colonies)
- -4.1-4.3 -> L4: Backbone pSB3K3 + Blue Promoter BBa_K238013 + GFP BBa_K145015 (74 min) (3 colonies)
- Primers used were Preffix FWD and Suffix REV, reactives needed for one reactions are as follows:
- PCR with Taq DNA polymerase
- Reactive (ul x sample)
- Taq Polymerase -> 1
- Taq Reaction Buffer 10X -> 5
- MgCl 50mM (can be used up to 3ul) -> 2.5
- dNTP’s 0.4ug/ul -> 2.5
- Primer Forward (can be used up to 3ul) -> 2.5
- Primer Reverse (can be used up to 3ul) -> 2.5
- HPLC -> 24
- DNA -> 10
- Total volume -> 50
- 1. 95ºC 5 min
- 2. 30 cycles
- 95ºC 45 seg
- 55ºC 45 seg
- 72ºC 1.5 min
- 3. 72ºC 10 min
- 4. Hold 4ºC
2. Run gel to verify PCR made on August 13th and 18th.
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