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Team:Paris Saclay/Notebook/August/1
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* Well 1 : 6µL of DNA Ladder | * Well 1 : 6µL of DNA Ladder | ||
Revision as of 23:31, 1 October 2013
Notebook : August 1
Lab work
A - Aerobic/Anaerobic regulation system / B - PCB sensing system
Objective : obtaining FNR and BphR2 proteins
1 - Gel purification of PCR products : BphR2 Part I, BphR2 Part II, FNR Part I, FNR Part II, RBS-FNR Part I PSB1C3
Xavier
Protocol : [http://www.mn-net.com/tabid/1452/default.aspx Gel purification ]
Nanodrop :
- BphR2 Part I : 44 ng/µL
- BphR2 Part II : 64 ng/µL
- FNR Part I : 147 ng/µL
- FNR Part II : 140 ng/µL
- RBS-FNR Part I : 167 ng/µL
- PSB1C3 : 159 ng/µL
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The PCR was good. Now we will do the Gibson assembly. |
2 - Gibson assembly.
Abdou, Xiaojing
Used quantities :
- BphR2 :
- PSB1C3 : 2µL
- BphR2 Part I : 1µL
- BphR2 Part II : 1µL
- Gibson mix : 15µL
- H2O : 1µL
- FNR :
- PSB1C3 : 2µL
- FNR Part I : 1µL
- FNR Part II : 1µL
- Gisbon mix : 15µL
- H2O : 1µL
- RBS-FNR :
- PSB1C3 : 2µL
- RBS-FNR Part I : 1µL
- FNR Part II : 1µL
- Gibson mix : 15µL
- H2O : 1µL
We let these mix at 50°C during 1h.
3 - PCR of : RBS-BphR2 Part I
Abdou
Used quantities :
- Oligo 54F : 1µL
- Oligo 55R : 1µL
- Buffer phusion : 5µL
- DNA : 0.25µL
- dNTP : 1µL
- Phusion : 5µL
- H2O : 36.5µL
PCR Program :
3 - Electrophoresis of the PCR products : RBS-BphR2 Part I
Xavier, XiaoJing
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Expected sizes :
- RBS-BphR2 Part I : 197pb
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We obtain fragments at the right size. We will purify it. |
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