Team:Paris Saclay/Notebook/August/19

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We spent our afternoon with Evry team. Each team presented its lab, modeling and human practices project. We learned informations to improve our human practices project. We discussed about safety and gave them some advices about the pills they would make.  
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Revision as of 15:33, 4 October 2013

Navigation Home Team Project Labwork Notebook Human practices

Contents

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

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

PSgel11908.jpg
  • Well 1 : 6µL DNA Ladder
  • Well 2 : 40µL BphR2 part I + 8µl of 6X loading dye
  • Gel : 1%

Expected size :

  • BphR2 Part I : 178 kb

We obtain a frangment at the right size. We can purify it.

2 - Gel purification of PCR products : BphR2 Part I

Damir

Protocol : Gel purification

Nanodrop :

  • BphR2 Part I: 57.2ng/µL

We lost our fragment BphR2 Part I. We will make the PCR again.

Lab work, Modeling, Human practices

Meeting with Evry

PSevry2.jpg

We spent our afternoon with Evry team. Each team presented its lab, modeling and human practices project. We learned informations to improve our human practices project. We discussed about safety and gave them some advices about the pills they would make.

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