Team:Paris Saclay/Notebook/August/12

From 2013.igem.org

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Anaïs, Nadia, XiaoJing
Anaïs, Nadia, XiaoJing
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Protocol : [[Team:Paris_Saclay/Protocols/digestion|digestion]]
 
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Used quantities :
 
* Pfnr in PSB1C3 (Bba_K1155000) :  
* Pfnr in PSB1C3 (Bba_K1155000) :  
** Buffer FD : 5µL
** Buffer FD : 5µL
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*PSB1C3 :  
*PSB1C3 :  
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  We can't see any band for Bba_K1155000 digestion. We just do it again and let it in the fridge over night.  
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  We can't see any band for Bba_K1155000 digestion. We just do it again and let it in the fridge over night. We obtain RBS-LacZ-Term and RBS-AmilCP-Term fragments. the digestion was good. We can now purify it.
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...
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==='''A - Aerobic/Anaerobic regulation system / B - PCB sensor system'''===
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Objective : obtaining FNR and BphR2 proteins
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===='''1 - Electrophoresis of PCR products : RBS-BphR2 part I, BphR2 part I, BphR2 part II, RBS-FNR part I, FNR part I and FNR part II '''====
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Damir
{{Team:Paris_Saclay/incl_fin}}
{{Team:Paris_Saclay/incl_fin}}

Revision as of 09:49, 20 August 2013

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Contents

Notebook : August 12

Lab work

A - Aerobic/Anaerobic regulation system

Obtaining Pfnr_RBS-LacZ-Term in PSB1C3

1 - Digestion of Bba_K1155000, Bba_K1155007 and Bba_K1155003

Anaïs, Nadia, XiaoJing

  • Pfnr in PSB1C3 (Bba_K1155000) :
    • Buffer FD : 5µL
    • H2O : 38µL
    • Pfnr : 5µL
    • SpeI : 1µL
    • PstI : 1µL
  • RBS-LacZ-Term in PSB1C3 (Bba_K1155007) :
    • Buffer FD : 5µL
    • H2O : 23µL
    • RBS-LacZ-Term : 20µL
    • XBal : 1µL
    • PstI : 1µL
  • RBS-AmilCP-Term in PSB1C3 (Bba_K1155003) :
    • Buffer FD : 5µL
    • H2O : 33µL
    • RBS-AmilCP-Term : 10µL
    • XBal : 1µL
    • PstI : 1µL

2 - Electrophoresis of digestion pieces

We gonna check if the digestion was good by control the size of digestion pieces.

Psdigestion12.jpg
  • Well 1 : 6µL DNA Ladder
  • Well 2 : 5µL of digested Bba_K1155000+1µl of 6X loading dy
  • Well 3 : 5µL of digested Bba_K1155007+1µl of 6X loading dy
  • Well 4 : 5µL of digested Bba_K1155003+1µl of 6X loading dy
  • Gel : 0.8%

Expected sizes :

  • RBS_LacZ_Term : 3500 kb
  • RBS_AmilCP_Term :
  • PSB1C3 : 
We can't see any band for Bba_K1155000 digestion. We just do it again and let it in the fridge over night. We obtain RBS-LacZ-Term and RBS-AmilCP-Term fragments. the digestion was good. We can now purify it.

A - Aerobic/Anaerobic regulation system / B - PCB sensor system

Objective : obtaining FNR and BphR2 proteins

1 - Electrophoresis of PCR products : RBS-BphR2 part I, BphR2 part I, BphR2 part II, RBS-FNR part I, FNR part I and FNR part II

Damir


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