Team:Paris Saclay/Notebook/August/19

From 2013.igem.org

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==='''A - Aerobic/Anaerobic regulation system'''===
==='''A - Aerobic/Anaerobic regulation system'''===
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===='''Objective : obtaining BBa_K1155007'''====
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====1 - Sequence analysis of BBa_K1155007====
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N'Guyen
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{|
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Our sequence analysis was good. We obtain our biobrick BBa_K1155007.
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===='''Objective : characterize BBa_K1155000, BBa_K1155004, BBa_K155005, BBa_K1155006 '''====
===='''Objective : characterize BBa_K1155000, BBa_K1155004, BBa_K155005, BBa_K1155006 '''====
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====2 - Ligation of NarK, NarG or NirB with RBS-LacZ-Term or RBS-AmilCP-Term in PSB1C3====
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====2 - Ligation of NarK, NarG or NirB with RBS-LacZ-Term or RBS-AmilCP-Term in pSB1C3====
XiaoJing  
XiaoJing  
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** Ligase : 1µL
** Ligase : 1µL
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====3 - Transformation of NarK, NarG or NirB with RBS-LacZ-Term or RBS-AmilCP-Term in PSB1C3 in DH5α====
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====3 - Transformation of NarK, NarG or NirB with RBS-LacZ-Term or RBS-AmilCP-Term in pSB1C3 in DH5α====
XiaoJing
XiaoJing
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Protocol : [[Team:Paris_Saclay/Protocols/Transformation|Bacterial transformation]]
Protocol : [[Team:Paris_Saclay/Protocols/Transformation|Bacterial transformation]]
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====4- Transformation of MG1655Z1 Δfnr::Km====
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====4- Make competent cells of strain MG1655Z1 Δfnr::Km====
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XiaoJing
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Protocol : [[Team:Paris_Saclay/preparation|Preparation of super competent cells]]
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====5- Transformation of MG1655Z1 Δfnr::Km====
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XiaoJing
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XioaJing
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We transformed pcp20 plasmid into strain MG1655Z1 Δfnr::Km. Then we spread in LB and ampicillin antibiotic plate. We need to incubate it at 30°C for two days.
Protocol : [[Team:Paris_Saclay/Protocols/Transformation|Bacterial transformation]]
Protocol : [[Team:Paris_Saclay/Protocols/Transformation|Bacterial transformation]]
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*Well 1 : 6µL DNA Ladder
*Well 1 : 6µL DNA Ladder
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*Well 2 : 40µL BphR2 part I+8µl of 6X loading dye
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*Well 2 : 40µL BphR2 part I + 8µl of 6X loading dye
*Gel : 1%
*Gel : 1%
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We obtain a frangment at the right size. We can purify it.
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We obtained a frangment at the right size. We can purify it.
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[[File:PSevry2.jpg]]
[[File:PSevry2.jpg]]
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We spent our afternoon with Evry team. Each team presented its lab, modeling and human practise projet. We argued about safety and gave them some advices about the pills they would make.
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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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Latest revision as of 01:29, 5 October 2013

Navigation Home Team Project Labwork Notebook Human practices

Contents

Notebook : August 19

Lab work

A - Aerobic/Anaerobic regulation system

Objective : obtaining BBa_K1155007

1 - Sequence analysis of BBa_K1155007

N'Guyen

Our sequence analysis was good. We obtain our biobrick BBa_K1155007.

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- Make competent cells of strain MG1655Z1 Δfnr::Km

XiaoJing

Protocol : Preparation of super competent cells

5- Transformation of MG1655Z1 Δfnr::Km

XiaoJing

We transformed pcp20 plasmid into strain MG1655Z1 Δfnr::Km. Then we spread in LB and ampicillin antibiotic plate. We need to incubate it at 30°C for two days.

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 obtained 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

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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