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Team:Paris Saclay/Notebook/September/9
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Contents |
Lab Work
By Mélanie
D-Lemon scent
extraction of TOPO plasmide with PS
Extraction with a Dna Kit extraction
[photo]
I failed
PCR
Transformation show some clone so I do a PCR.
| 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:
Pu Pr (universal primer)
| 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 1 GS (8 clones) ans cad (8 clones) photo 2 LS (8 clones)
After these results I choose some clones to do a liquid culture
