Team:Paris Saclay/Notebook/July/23

From 2013.igem.org

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===='''Objective : obtaining Bba_K1155003, Bba_K1155007'''====
===='''Objective : obtaining Bba_K1155003, Bba_K1155007'''====
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===='''1 - Electrophoresis of the digestion of Bba_I732017 by EcoRI/SpeI,  Bba_K592009 SpeI/PstI, Bba_B0015, Bba_B0017 by XbaI/PstI '''====
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===='''1 - Electrophoresis of the digestion of Bba_I732017 by EcoRI/SpeI,  Bba_K592009 by SpeI/PstI, Bba_B0015, Bba_B0017 by XbaI/PstI '''====
Abdou, Anaïs, Xavier, Zhou
Abdou, Anaïs, Xavier, Zhou
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|}
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===='''6 - Culture of Bba_I732017 and Bba_K592009'''====
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===='''6 - Digestion of Bba_K592009 by SpeI/PstI'''====
 +
 
 +
Anaïs
 +
 
 +
Used quantities :
 +
* Buffer FD : 3µL
 +
* DNA : 15µL
 +
* SpeI FD : 2µL
 +
* PstI : 2µL
 +
* H2O : 8µL
 +
 
 +
===='''7 - Electrophoresis to check the digestion of Bba_K592009 by SpeI/PstI'''====
 +
 
 +
Zhou
 +
 
 +
{|
 +
| style="width:350px;border:1px solid black;" |[[]]
 +
| style="width:350px;border:1px solid black;vertical-align:top;" |
 +
* Well 1 : 6µL of DNA 6X loading dye
 +
* Well 2 to 4 : 5µL of Bba_K592009 digested by SpeI/PstI+1µL of 6X loading dye
 +
* Gel : 0.8%
 +
|}
 +
 
 +
Expected sizes :
 +
* Amil CP :
 +
 
 +
{|
 +
| style="border:1px solid black;padding:5px;background-color:#DEDEDE;" |
 +
We can't see anything. The purification wasn't good. We will do cultures again.
 +
|}
 +
 
 +
===='''8 - Culture of Bba_I732017 and Bba_K592009'''====
Abdou, Xavier
Abdou, Xavier

Revision as of 21:45, 26 September 2013

Navigation Home Team Project Labwork Notebook Human practices

Contents

Notebook : July 23

Lab work

A - Aerobic/Anaerobic regulation system

Objective : obtaining Bba_K1155003, Bba_K1155007

1 - Electrophoresis of the digestion of Bba_I732017 by EcoRI/SpeI, Bba_K592009 by SpeI/PstI, Bba_B0015, Bba_B0017 by XbaI/PstI

Abdou, Anaïs, Xavier, Zhou

[[]]
  • Well 1 to 3: 30µL of Bba_I732017 digested by EcoRI/SpeI+6µL of 6X loading dye
  • Well 4 : 6µL of DNA 6X loading dye
  • Gel : 0.8%
[[]]
  • Well 1 : 5µL of Bba_B0015 digested by XbaI/PstI+1µL of 6X loading dye
  • Well 2 : 5µL of Bba_B0017 digested by XbaI/PstI+1µL of 6X loading dye
  • Well 3 : 5µL of Bba_K592009 digested SpeI/PstI+1µL of 6X loading dye
  • Well 4 : 6µL of DNA 6X loading dye
  • Gel : 0.8%

Expected sizes :

  • RBS-LacZ :
  • Amil CP :
  • Term :
  • PSB3A1 :

We obtain a RBS-lacZ and Term fragments at the right size. The digestions were good and we will purify them.

2 - Gel purification of digestion products : RBS-LacZ

Xavier

Protocol : [http://www.mn-net.com/tabid/1452/default.aspx Gel purification ]

3 - Electrophoresis to check the gel purification of digestion products : RBS-LacZ

Abdou

[[]]
  • Well 1 : 6µL of DNA 6X loading dye
  • Well 2: 5µL of RBS-LacZ+1µL of 6X loading dye
  • Gel : 0.8%

Expected sizes :

  • RBS-LacZ :

We can't see anything. The purification wasn't good. We will do the digestion again.

4 - Digestion of Bba_I732017 by EcoRI/PstI

Anaïs

Used quantities :

  • Bba_I732017 : 10µL
  • Buffer FD : 3µL
  • EcoRI FD : 1.5µL
  • PstI FD : 1.5µL
  • H2O : 14µL

5 - Electrophoresis to check the gel digestion of Bba_I732017 by EcoRI/SpeI

Zhou

We can't see anything. The digestion wasn't good, we will do it the again.

6 - Digestion of Bba_K592009 by SpeI/PstI

Anaïs

Used quantities :

  • Buffer FD : 3µL
  • DNA : 15µL
  • SpeI FD : 2µL
  • PstI : 2µL
  • H2O : 8µL

7 - Electrophoresis to check the digestion of Bba_K592009 by SpeI/PstI

Zhou

[[]]
  • Well 1 : 6µL of DNA 6X loading dye
  • Well 2 to 4 : 5µL of Bba_K592009 digested by SpeI/PstI+1µL of 6X loading dye
  • Gel : 0.8%

Expected sizes :

  • Amil CP :

We can't see anything. The purification wasn't good. We will do cultures again.

8 - Culture of Bba_I732017 and Bba_K592009

Abdou, Xavier


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