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Team:Paris Saclay/Notebook/September/4
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I use the same PCR condition than [https://2014.igem.org/Team:Paris_Saclay/Notebook/September/2#Limonene_synthase September 2] but with the right primer and plasmid | I use the same PCR condition than [https://2014.igem.org/Team:Paris_Saclay/Notebook/September/2#Limonene_synthase September 2] but with the right primer and plasmid | ||
| - | [ | + | [[File:0309 Vérif PCR GS CAD PS LS Chromo.jpg|400px]] |
We can see that we don't have any insert in our plasmid | We can see that we don't have any insert in our plasmid | ||
Revision as of 12:06, 7 September 2014
Contents |
Lab Work
D- Lemon Scent
By Mélanie
Verfication of the pPS3/4/5 plasmid
Due to the strange results obtained last day, I do a PCR of the insert with the differents plasmid : I use the same PCR condition than September 2 but with the right primer and plasmid
We can see that we don't have any insert in our plasmid
PCR verification
We find some other PCR of BBa K762100 and GS and BBa_K517003 so we check it by electrophoresis
And I purify the PCR
Well 1 = Ladder
Well 2 = GS
Well 3 = PS
Well 4 = CAD
We can see that we don't have lost a lot of our PCR product
cloning
Cloning of PCR purification in a TA plasmid First I had to add AAA to the PCR
| component | volume |
|---|---|
| buffer | 1μl |
| dATP | 1μl |
| PCR | 7.5μl |
| Taq | 0.5μl |
72° 15'
and
| component | volume |
|---|---|
| H2O | 1μl |
| buffer | 1μl |
| vector | 1μl |
| cloning | 1μl |
30' at room temperature
and transformation with competent bacteria add DNA 30' on ice 45' 42°c 30' at 37°
spread on petri dish
