Team:Paris Saclay/Notebook/September/9
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- | [[File:0909 PCR CLONE LS.jpg| | + | |+ |
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- | + | | scope=col | [[File:0909 PCR CLONE.jpg|330px|center]] | |
- | + | | scope=col | [[File:0909 PCR CLONE LS.jpg|330px|center]] | |
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+ | | GS (8 clones) ans cad (8 clones) | ||
+ | | LS (8 clones) | ||
+ | |} | ||
After these results I choose some clones to do a liquid culture | After these results I choose some clones to do a liquid culture |
Latest revision as of 13:00, 14 October 2014
Contents |
Tuesday 9th september
Lab Work
By Mélanie
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 |
GS (8 clones) ans cad (8 clones) | LS (8 clones) |
After these results I choose some clones to do a liquid culture