Team:Paris Saclay/Notebook/August/12

From 2013.igem.org

(Difference between revisions)
Line 16: Line 16:
** Buffer FD : 5µL
** Buffer FD : 5µL
** H2O : 38µL
** H2O : 38µL
-
** Pfnr : 5µL
+
** DNA : 5µL
** SpeI : 1µL
** SpeI : 1µL
** PstI : 1µL
** PstI : 1µL
Line 23: Line 23:
** Buffer FD : 5µL  
** Buffer FD : 5µL  
** H2O : 23µL
** H2O : 23µL
-
** RBS-LacZ-Term : 20µL  
+
** DNA : 20µL  
** XBal : 1µL
** XBal : 1µL
** PstI : 1µL
** PstI : 1µL
Line 30: Line 30:
** Buffer FD : 5µL
** Buffer FD : 5µL
** H2O : 33µL
** H2O : 33µL
-
** RBS-AmilCP-Term : 10µL  
+
** DNA : 10µL  
** XBal : 1µL
** XBal : 1µL
** PstI : 1µL
** PstI : 1µL
===='''2 - Electrophoresis of digestion of Bba_K1155000 by SpeI and PstI, Bba_K1155007 and Bba_K1155003 by XBalI and PstI'''====  
===='''2 - Electrophoresis of digestion of Bba_K1155000 by SpeI and PstI, Bba_K1155007 and Bba_K1155003 by XBalI and PstI'''====  
 +
 +
Nadia
{|
{|
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| style="width:350px;border:1px solid black;vertical-align:top;" |
| style="width:350px;border:1px solid black;vertical-align:top;" |
*Well 1 : 6µL DNA Ladder
*Well 1 : 6µL DNA Ladder
-
*Well 2 : 5µL of digested Bba_K1155000+1µl of 6X loading dy
+
*Well 2 : 5µL DNA+1µl of 6X loading dy
-
*Well 3 : 5µL of digested Bba_K1155007+1µl of 6X loading dy
+
*Well 3 : 5µL DNA+1µl of 6X loading dy
-
*Well 4 : 5µL of digested Bba_K1155003+1µl of 6X loading dy
+
*Well 4 : 5µL DNA+1µl of 6X loading dy
*Gel : 0.8%
*Gel : 0.8%
|}
|}
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Expected sizes :  
Expected sizes :  
*RBS_LacZ_Term : 3500 kb
*RBS_LacZ_Term : 3500 kb
-
*RBS_AmilCP_Term :  
+
*RBS_AmilCP_Term : ...
-
*PSB1C3 :  
+
*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.
+
  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'''====
===='''3 - Digestion of Bba_K1155000 by SpeI and PstI'''====
Anaïs, Nadia
Anaïs, Nadia
-
 
+
-
* Bba_K1155000 :
+
* Buffer FD : 5µL
-
** Buffer FD : 5µL
+
* H2O : 38µL
-
** H2O : 38µL
+
* DNA : 5µL
-
** Pfnr : 5µL
+
* SpeI : 1µL
-
** SpeI : 1µL
+
* PstI : 1µL
-
** PstI : 1µL
+
==='''A - Aerobic/Anaerobic regulation system / B - PCB sensor system'''===
==='''A - Aerobic/Anaerobic regulation system / B - PCB sensor system'''===
Line 72: Line 73:
Damir
Damir
-
 
+
{|
 +
| style="width:350px;border:1px solid black;" |[[]]
 +
| style="width:350px;border:1px solid black;vertical-align:top;" |
 +
*Well 1 : 6µL DNA Ladder
 +
*Well 2 : 5µL DNA+1µl of 6X loading dy
 +
*Well 3 : 5µL DNA+1µl of 6X loading dy
 +
*Well 4 : 5µL DNA+1µl of 6X loading dy
 +
*Gel : 0.8%
 +
|}
{{Team:Paris_Saclay/incl_fin}}
{{Team:Paris_Saclay/incl_fin}}

Revision as of 10:01, 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 XBaI and PstI

Anaïs, Nadia, XiaoJing

  • Bba_K1155000 :
    • Buffer FD : 5µL
    • H2O : 38µL
    • DNA : 5µL
    • SpeI : 1µL
    • PstI : 1µL
  • Bba_K1155007 :
    • Buffer FD : 5µL
    • H2O : 23µL
    • DNA : 20µL
    • XBal : 1µL
    • PstI : 1µL
  • Bba_K1155003 :
    • Buffer FD : 5µL
    • H2O : 33µL
    • DNA : 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

Nadia

Psdigestion12.jpg
  • Well 1 : 6µL DNA Ladder
  • Well 2 : 5µL DNA+1µl of 6X loading dy
  • Well 3 : 5µL DNA+1µl of 6X loading dy
  • Well 4 : 5µL DNA+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

  • Buffer FD : 5µL
  • H2O : 38µL
  • DNA : 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

[[]]
  • Well 1 : 6µL DNA Ladder
  • Well 2 : 5µL DNA+1µl of 6X loading dy
  • Well 3 : 5µL DNA+1µl of 6X loading dy
  • Well 4 : 5µL DNA+1µl of 6X loading dy
  • Gel : 0.8%

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