Team:Paris Saclay/Notebook/July/8

From 2013.igem.org

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==='''A - Aerobic/Anaerobic regulation system'''===
==='''A - Aerobic/Anaerobic regulation system'''===
-
===='''Objective : Bba_K1155003, Bba_K1155007'''====
+
===='''Objective : BBa_K1155003, BBa_K1155007'''====
-
===='''1 - Digestion of Bba_K592009, Bba_I732017 by NotI, XhoI, EcoRI, EcoRI/PstI'''====
+
===='''1 - Digestion of BBa_K592009, BBa_I732017 by NotI, XhoI, EcoRI, EcoRI/PstI'''====
Abou, Anaïs, Sheng, Zhou
Abou, Anaïs, Sheng, Zhou
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* NotI :  
* NotI :  
-
** Bba_K592009 : 2µL
+
** BBa_K592009 : 2µL
** Buffer orange : 2µL
** Buffer orange : 2µL
** NotI : 0.5µL
** NotI : 0.5µL
Line 22: Line 22:
* XhoI :  
* XhoI :  
-
** Bba_K592009 : 2µL
+
** BBa_K592009 : 2µL
** Buffer red : 2µL
** Buffer red : 2µL
** XhoI : 0.5µL
** XhoI : 0.5µL
Line 28: Line 28:
* EcoRI :  
* EcoRI :  
-
** Bba_K592009 : 2µL
+
** BBa_K592009 : 2µL
** Buffer orange : 2µL
** Buffer orange : 2µL
** EcoRI : 0.5µL
** EcoRI : 0.5µL
Line 34: Line 34:
* EcoRI/PstI :  
* EcoRI/PstI :  
-
** Bba_I732017 : 2µL
+
** BBa_I732017 : 2µL
** Buffer orange : 2µL
** Buffer orange : 2µL
** EcoRI : 0.5µL
** EcoRI : 0.5µL
Line 42: Line 42:
We let our digestion 1h30 at 37°C.
We let our digestion 1h30 at 37°C.
-
===='''2 - Electrophoresis of the digestion of Bba_K592009, Bba_I732017 by NotI, XhoI, EcoRI, EcoRI/PstI'''====
+
===='''2 - Electrophoresis of the digestion of BBa_K592009, BBa_I732017 by NotI, XhoI, EcoRI, EcoRI/PstI'''====
Abdou, Anaïs, Sheng, Zhou
Abdou, Anaïs, Sheng, Zhou
{|
{|
-
| style="width:350px;border:1px solid black;" |[[File:Psgel0807.jpg|300px]]
+
| style="width:350px;border:1px solid black;" |[[File:Psgel0807.jpg|500px]]
| 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 Bba_K592009 clone 1 digested by EcoRI+1µL of 6X loading dye
+
* Well 2 : 5µL BBa_K592009 clone 1 digested by EcoRI+1µL of 6X loading dye
-
* Well 3 : 5µL Bba_K592009 clone 2 digested by EcoRI+1µL of 6X loading dye
+
* Well 3 : 5µL BBa_K592009 clone 2 digested by EcoRI+1µL of 6X loading dye
-
* Well 4 : 5µL Bba_K592009 clone 1 digested by NotI+1µL of 6X loading dye
+
* Well 4 : 5µL BBa_K592009 clone 1 digested by NotI+1µL of 6X loading dye
-
* Well 5 : 5µL Bba_K592009 clone 2 digested by NotI+1µL of 6X loading dye
+
* Well 5 : 5µL BBa_K592009 clone 2 digested by NotI+1µL of 6X loading dye
-
* Well 6 : 5µL Bba_K592009 clone 1 digested by XhoI+1µL of 6X loading dye
+
* Well 6 : 5µL BBa_K592009 clone 1 digested by XhoI+1µL of 6X loading dye
-
* Well 7 : 5µL Bba_K592009 clone 2 digested by XhoI+1µL of 6X loading dye
+
* Well 7 : 5µL BBa_K592009 clone 2 digested by XhoI+1µL of 6X loading dye
-
* Well 8 : 5µL Bba_I732017 clone 1 digested by EcoRI/PstI+1µL of 6X loading dye
+
* Well 8 : 5µL BBa_I732017 clone 1 digested by EcoRI/PstI+1µL of 6X loading dye
-
* Well 9 : 5µL Bba_I732017 clone 2 digested by EcoRI/PstI+1µL of 6X loading dye
+
* Well 9 : 5µL BBa_I732017 clone 2 digested by EcoRI/PstI+1µL of 6X loading dye
* Well 10 : -
* Well 10 : -
* Well 11 : -
* Well 11 : -
Line 65: Line 65:
Expected sizes :  
Expected sizes :  
-
* Bba_K592009 digested by NotI : 2046bp + 693bp
+
* BBa_K592009 digested by NotI : 2046bp + 693bp
-
* Bba_K592009 digested by XhoI : 1842bp + 892bp
+
* BBa_K592009 digested by XhoI : 1842bp + 892bp
-
* Bba_I732017 : ...
+
* BBa_I732017 : 3093bp
{|
{|

Latest revision as of 23:55, 4 October 2013

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Contents

Notebook : July 8

Lab work

A - Aerobic/Anaerobic regulation system

Objective : BBa_K1155003, BBa_K1155007

1 - Digestion of BBa_K592009, BBa_I732017 by NotI, XhoI, EcoRI, EcoRI/PstI

Abou, Anaïs, Sheng, Zhou

Used quantities :

  • NotI :
    • BBa_K592009 : 2µL
    • Buffer orange : 2µL
    • NotI : 0.5µL
    • H2O : 15.5µL
  • XhoI :
    • BBa_K592009 : 2µL
    • Buffer red : 2µL
    • XhoI : 0.5µL
    • H2O : 15.5µL
  • EcoRI :
    • BBa_K592009 : 2µL
    • Buffer orange : 2µL
    • EcoRI : 0.5µL
    • H2O : 15.5µL
  • EcoRI/PstI :
    • BBa_I732017 : 2µL
    • Buffer orange : 2µL
    • EcoRI : 0.5µL
    • PstI : 0.5µL
    • H2O : 15µL

We let our digestion 1h30 at 37°C.

2 - Electrophoresis of the digestion of BBa_K592009, BBa_I732017 by NotI, XhoI, EcoRI, EcoRI/PstI

Abdou, Anaïs, Sheng, Zhou

Psgel0807.jpg
  • Well 1 : 6µL DNA Ladder
  • Well 2 : 5µL BBa_K592009 clone 1 digested by EcoRI+1µL of 6X loading dye
  • Well 3 : 5µL BBa_K592009 clone 2 digested by EcoRI+1µL of 6X loading dye
  • Well 4 : 5µL BBa_K592009 clone 1 digested by NotI+1µL of 6X loading dye
  • Well 5 : 5µL BBa_K592009 clone 2 digested by NotI+1µL of 6X loading dye
  • Well 6 : 5µL BBa_K592009 clone 1 digested by XhoI+1µL of 6X loading dye
  • Well 7 : 5µL BBa_K592009 clone 2 digested by XhoI+1µL of 6X loading dye
  • Well 8 : 5µL BBa_I732017 clone 1 digested by EcoRI/PstI+1µL of 6X loading dye
  • Well 9 : 5µL BBa_I732017 clone 2 digested by EcoRI/PstI+1µL of 6X loading dye
  • Well 10 : -
  • Well 11 : -
  • Well 12 : 6µL DNA Ladder
  • Gel : 1%

Expected sizes :

  • BBa_K592009 digested by NotI : 2046bp + 693bp
  • BBa_K592009 digested by XhoI : 1842bp + 892bp
  • BBa_I732017 : 3093bp

We obtain fragments at the right size. Our extraction was good.


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