Team:Paris/August 26
From 2008.igem.org
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{{Paris/Calendar_Links|August 25|August 27}} | {{Paris/Calendar_Links|August 25|August 27}} | ||
- | = | + | =PCR1= |
* pFlhDC (O111-F / O113-R) | * pFlhDC (O111-F / O113-R) | ||
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O2=1µL<br> | O2=1µL<br> | ||
Phusion=0,2µL<br> | Phusion=0,2µL<br> | ||
+ | |||
+ | =PCR2= | ||
+ | |||
+ | * pFliA (rbs) (O145-F / O144-R) | ||
+ | * pFliA (O145-F / O146-R) | ||
+ | * pFliA +Gene FliA (O145-F / O151-R) | ||
+ | ---- | ||
+ | |||
+ | '''Program: promoter'''<br> | ||
+ | '''''Denaturation :''''' <br> | ||
+ | 98°C-5'<br> | ||
+ | '''''Cycling 1 (3X) :''''' <br> | ||
+ | 98°C-10"<br> | ||
+ | 55°C - 30''<br> | ||
+ | 72°C-30"<br> | ||
+ | '''''Cycling 2 (30X) :''''' <br> | ||
+ | 98°C-10"<br> | ||
+ | 65°C-30" | ||
+ | 72°C-30"<br> | ||
+ | '''''Elongation :''''' <br> | ||
+ | 72°C-5'<br> | ||
+ | |||
+ | ---- | ||
+ | |||
+ | Vf=50µL<br> | ||
+ | H20=33,5µL<br> | ||
+ | Buffer5X=10µL<br> | ||
+ | dNTP=1µL<br> | ||
+ | O1=2,5µL<br> | ||
+ | O2=2,5µL<br> | ||
+ | Phusion=0,5µL<br> | ||
+ | |||
+ | =PCR3= | ||
+ | |||
+ | * PCRFliA1 (O143-F / O152-R) | ||
+ | * PCRFliA2 (O153-F / O150-R) | ||
+ | * PCRFliA3 (O148-F / O150-R) | ||
+ | |||
+ | ---- | ||
+ | |||
+ | '''Program: PCRFliA1'''<br> | ||
+ | '''''Denaturation :''''' <br> | ||
+ | 98°C-5'<br> | ||
+ | '''''Cycling 1 (30X) :''''' <br> | ||
+ | 98°C-10"<br> | ||
+ | Gradient 66°C +/-6°C - 25''<br> | ||
+ | 72°C-20"<br> | ||
+ | '''''Elongation :''''' <br> | ||
+ | 72°C-5'<br> | ||
+ | |||
+ | '''Program: PCRFliA2'''<br> | ||
+ | '''''Denaturation :''''' <br> | ||
+ | 98°C-5'<br> | ||
+ | '''''Cycling 1 (3X) :''''' <br> | ||
+ | 98°C-10"<br> | ||
+ | Gradient 61°C +/-10°C - 25''<br> | ||
+ | 72°C-20"<br> | ||
+ | '''''Cycling 2 (30X) :''''' <br> | ||
+ | 98°C-10"<br> | ||
+ | 72°C-30"<br> | ||
+ | '''''Elongation :''''' <br> | ||
+ | 72°C-5'<br> | ||
=Cloning of EnvZ*= | =Cloning of EnvZ*= |
Revision as of 11:05, 26 August 2008
PCR1
Program: Gradient Vf=20µL PCR2
Program: promoter Vf=50µL PCR3
Program: PCRFliA1 Program: PCRFliA2 Cloning of EnvZ*Concentration measurement by Biophotometer
Ligation |