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Team:Paris Saclay/Notebook/August/19
From 2013.igem.org
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====2 - Gel purification of PCR products : BphR2 Part I ==== | ====2 - Gel purification of PCR products : BphR2 Part I ==== | ||
Revision as of 14:14, 2 October 2013
Notebook : August 19
Lab work
A - Aerobic/Anaerobic regulation system
Objective : characterize BBa_K1155000, BBa_K1155004, BBa_K155005, BBa_K1155006
1 - EtOH precipitation of RBS-LacZ-Term and RBS-AmilCP-Term
Nadia
Protocol : EtOH precipitation
We used 20µL of DNA.
Nanodrop :
- RBS-LacZ-Term : 38.4ng/µL
- RBS-AmilCP-Term : 28.7ng/µL
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The precipitation was good. We will make ligations. |
2 - Ligation of NarK, NarG or NirB with RBS-LacZ-Term or RBS-AmilCP-Term in PSB1C3
XiaoJing
Used quantities :
- NirB and RBS-LacZ-Term :
- NirB : 3µL
- RBS-LacZ-Term : 1µL
- Buffer : 2µL
- H2O : 13µL
- Ligase : 1µL
- NirB and RBS-AmilCP-Term :
- NirB : 3µL
- RBS-AmilCP-Term : 1µL
- Buffer : 2µL
- H2O : 13µL
- Ligase : 1µL
- NarG and RBS-LacZ-Term :
- NarG : 3µL
- RBS-LacZ-Term : 1µL
- Buffer : 2µL
- H2O : 13µL
- Ligase : 1µL
- NarG and RBS-AmilCP-Term :
- NarG : 3µL
- RBS-AmilCP-Term : 1µL
- Buffer : 2µL
- H2O : 13µL
- Ligase : 1µL
- NarK and RBS-LacZ-Term :
- Nar K : 3µL
- RBS-LacZ-Term : 1µL
- Buffer : 2µL
- H2O : 13µL
- Ligase : 1µL
- NarK and RBS-AmilCP-Term :
- NarK : 3µL
- RBS-AmilCP-Term : 1µL
- Buffer : 2µL
- H2O : 13µL
- Ligase : 1µL
3 - Transformation of NarK, NarG or NirB with RBS-LacZ-Term or RBS-AmilCP-Term in PSB1C3 in DH5α
XiaoJing
Protocol : Bacterial transformation
4- Transformation of MG1655Z1 Δfnr::Km
XioaJing
Protocol : Bacterial transformation
A - Aerobic/Anaerobic regulation system / B - PCB sensor system
Objective : obtaining FNR and BphR2 proteins
1 - Electrophoresis gel of PCR products : BphR2 Part I
Damir
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Expected size :
- BphR2 Part I : 178 kb
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We obtain a frangment at the right size. We can purify it. |
2 - Gel purification of PCR products : BphR2 Part I
Damir
Protocol : [http://www.mn-net.com/tabid/1452/default.aspx Gel purification ]
Nanodrop :
- BphR2 Part I: 57.2ng/µL
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We lost our fragment BphR2 Part I. We will make the PCR again. |
Lab work, Modeling, Human practices
Meeting with Evry
We spent our afternoon with Evry team. Each team presented its lab, modeling and human practise projet. ..........
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