Team:Paris Saclay/Notebook/August/12

From 2013.igem.org

(Difference between revisions)
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'''Obtaining Pfnr_RBS-LacZ-Term in PSB1C3'''
'''Obtaining Pfnr_RBS-LacZ-Term in PSB1C3'''
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===='''1 - Digestion of Bba_K1155000, Bba_K1155007 and Bba_K1155003'''====
+
===='''1 - Digestion of Bba_K1155000 by SpeI and PstI, Bba_K1155007 and Bba_K1155003 by XBalI and PstI'''====
Anaïs, Nadia, XiaoJing
Anaïs, Nadia, XiaoJing
-
* Pfnr in PSB1C3 (Bba_K1155000) :  
+
* Bba_K1155000 :  
** Buffer FD : 5µL
** Buffer FD : 5µL
** H2O : 38µL
** H2O : 38µL
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** PstI : 1µL
** PstI : 1µL
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* RBS-LacZ-Term in PSB1C3 (Bba_K1155007) :  
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* Bba_K1155007 :  
** Buffer FD : 5µL  
** Buffer FD : 5µL  
** H2O : 23µL
** H2O : 23µL
Line 27: Line 27:
** PstI : 1µL
** PstI : 1µL
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* RBS-AmilCP-Term in PSB1C3 (Bba_K1155003) :  
+
* Bba_K1155003 :  
** Buffer FD : 5µL
** Buffer FD : 5µL
** H2O : 33µL
** H2O : 33µL
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** PstI : 1µL
** PstI : 1µL
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===='''2 - Electrophoresis of digestion pieces'''====  
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===='''2 - Electrophoresis of digestion of Bba_K1155000 by SpeI and PstI, Bba_K1155007 and Bba_K1155003 by XBalI and PstI'''====  
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We gonna check if the digestion was good by control the size of digestion pieces.
 
{|
{|
| style="width:350px;border:1px solid black;" |[[File:Psdigestion12.jpg|350px]]
| style="width:350px;border:1px solid black;" |[[File:Psdigestion12.jpg|350px]]
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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. We obtain RBS-LacZ-Term and RBS-AmilCP-Term fragments. the digestion was good. We can now purify it.
+
  We can't see any band for Bba_K1155000 digestion. The digestion failed. We will do it again. We obtain RBS-LacZ-Term and RBS-AmilCP-Term fragments. the digestion was good. We can now purify it.
 +
 
 +
===='''3 - Digestion of Bba_K1155000 by SpeI and PstI'''====
 +
 
 +
Anaïs, Nadia
 +
 
 +
* Bba_K1155000 :
 +
** Buffer FD : 5µL
 +
** H2O : 38µL
 +
** Pfnr : 5µL
 +
** SpeI : 1µL
 +
** PstI : 1µL
==='''A - Aerobic/Anaerobic regulation system / B - PCB sensor system'''===
==='''A - Aerobic/Anaerobic regulation system / B - PCB sensor system'''===
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Objective : obtaining FNR and BphR2 proteins
+
'''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 '''====
===='''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
Damir
 +
{{Team:Paris_Saclay/incl_fin}}
{{Team:Paris_Saclay/incl_fin}}

Revision as of 09:56, 20 August 2013

Navigation Home Team Project Labwork Notebook Human practices

Contents

Notebook : August 12

Lab work

A - Aerobic/Anaerobic regulation system

Obtaining Pfnr_RBS-LacZ-Term in PSB1C3

1 - Digestion of Bba_K1155000 by SpeI and PstI, Bba_K1155007 and Bba_K1155003 by XBalI and PstI

Anaïs, Nadia, XiaoJing

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

2 - Electrophoresis of digestion of Bba_K1155000 by SpeI and PstI, Bba_K1155007 and Bba_K1155003 by XBalI and PstI

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. The digestion failed. We will do it again. We obtain RBS-LacZ-Term and RBS-AmilCP-Term fragments. the digestion was good. We can now purify it.

3 - Digestion of Bba_K1155000 by SpeI and PstI

Anaïs, Nadia

  • Bba_K1155000 :
    • Buffer FD : 5µL
    • H2O : 38µL
    • Pfnr : 5µL
    • SpeI : 1µL
    • PstI : 1µL

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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