Team:Paris Saclay/Notebook/September/8
From 2014.igem.org
Contents |
Lab Work
D-Lemon Scent
Checking PCR
Electrophoresis of the PCR made Friday
[photo]
PCR with bacteria
From the petri dish (made here, I select some clone to do a PCR : clones 1-2-3 for cad clones 4-5 for PS clones 6 for LS
component | volume |
---|---|
H2O | 41.5μl |
buffer | 5μl |
dNTPs | 1μl |
Primer 1 | 1μl |
Primer 2 | 1μl |
bacteria | (about 1µl = 1 colony) |
Dream taq | 0.5μl |
Primer used:
For cloning in Topo vector = Pu Pr (universal primer)
For pPS2 = iPS 66/67
Cycle step | Temperature | Time | Cycle |
---|---|---|---|
Bacteria lysis |
95°C |
5 min |
1 |
Denaturation | 94°C | 30 s | 25 |
Annealing | 50°C | 25 s | 25 |
Extension | 72°C | 1 min | 25 |
Final extension | 72°C | 10 min | 1 |
Final extension | 8°C | hold | 1 |
[photo]
We identify that we have success with to clone of PS.
Liquide culture of clones
The PS clone that reveal to be ok are cultivate in LB + AMP médium