Team:Paris Saclay/Notebook/July/5

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(→2 - Extraction of Bba_K1155000 and Bba_K592009 from DH5α)
(→1 - Electrophoresis of the digestion of BBa_K1155000 by NotI, HpaI, MluI)
 
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==='''A - Aerobic/Anaerobic regulation system'''===
==='''A - Aerobic/Anaerobic regulation system'''===
-
===='''Objective : obtaining Bba_K1155000'''====
+
===='''Objective : obtaining BBa_K1155000'''====
-
===='''1 - Electrophoresis of the digestion of Bba_K1155000 by NotI, HpaI, MluI'''====
+
===='''1 - Electrophoresis of the digestion of BBa_K1155000 by NotI, HpaI, MluI'''====
Sheng, Zhou
Sheng, Zhou
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| style="width:350px;border:1px solid black;" | [[File:PCRPS050813.jpg|right|350px]]
| style="width:350px;border:1px solid black;" | [[File:PCRPS050813.jpg|right|350px]]
| style="width:350px;border:1px solid black;" |
| style="width:350px;border:1px solid black;" |
-
* Well 1, 2 : 5µL of Bba_K1155000 digested by Not I+1µL of 6X loading dye
+
* Well 1, 2 : 5µL of BBa_K1155000 digested by Not I+1µL of 6X loading dye
-
* Well 3, 4 : 5µL of Bba_K1155000 digested by Hpa I+1µL of 6X loading dye
+
* Well 3, 4 : 5µL of BBa_K1155000 digested by Hpa I+1µL of 6X loading dye
* Well 5 : 6µL of DNA Ladder
* Well 5 : 6µL of DNA Ladder
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* Well 6, 7 : 5µL of Bba_K1155000 digested by Mlu I+1µL of 6X loading dye
+
* Well 6, 7 : 5µL of BBa_K1155000 digested by Mlu I+1µL of 6X loading dye
* Gel 0.8%
* Gel 0.8%
|}
|}
Expected sizes :  
Expected sizes :  
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* Bba_K1155000 : ...
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* BBa_K1155000 : 111bp
{|
{|
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|}
|}
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===='''2 - Extraction of Bba_K1155000 and Bba_K592009 from DH5α'''====
+
===='''2 - Extraction of BBa_K1155000 from DH5α'''====
Abdou
Abdou
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Transformation of 07/04 works. We will extract DNA plasmid.
Transformation of 07/04 works. We will extract DNA plasmid.
|}
|}
-
Protocol : [[Team:Paris_Saclay/extraction|Hight copy plamid extraction]]
+
 
 +
Protocol : [[Team:Paris_Saclay/extraction|High copy plamid extraction]]
Nanodrop :  
Nanodrop :  
-
* Bba_K1155000P : 761ng/µl
+
* BBa_K1155000 : 761ng/µl
-
* Bba_K592009 : 229ng/µl
+
{|
{|
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|}
|}
 +
===='''Objective : obtaining BBa_K1155003, BBa_K1155007'''====
 +
===='''1 - Extraction of BBa_K592009 and BBa_I732017 from DH5α'''====
-
<u>Electrophoresis band size estimation</u><br>
+
Abdou
-
<p>We used Clonemanager for band size estimation:</p>
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-
<br>
+
-
{| border="1" align="center"
+
-
|-
+
-
|Molecule
+
-
|enzymes(Buffer)
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-
|Size
+
-
|-
+
-
|AmilCP clone
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-
|Nod I(Orange)
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-
|2046+693bp
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-
|-
+
-
|AmilCP clone
+
-
|XHOI(Red)
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-
|1842+892bp
+
-
|-
+
-
|BBa_K1155000
+
-
|XHOI(Red)
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-
|1289+892bp
+
-
|-
+
-
|BBa_K1155000
+
-
|ECoR I(Orange)
+
-
|2151bp
+
-
|}<br>
+
-
 
+
-
 
+
-
 
+
-
 
+
-
 
+
-
 
+
-
 
+
-
 
+
-
 
+
-
<br>
+
 +
{|
 +
| style="border:1px solid black;padding:5px;background-color:#DE;" |
 +
Transformation of 07/04 works. We will extract DNA plasmid.
 +
|}
 +
Protocol : [[Team:Paris_Saclay/extraction|High copy plamid extraction]]
 +
Nanodrop :
 +
* BBa_K592009 : 229ng/µl
 +
{|
 +
| style="border:1px solid black;padding:5px;background-color:#DEDEDE;" |
 +
By using NanoDrop, we found some errors which could be due to device malfunctional issue.
 +
|}

Latest revision as of 16:35, 4 October 2013

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Contents

Notebook : July 5

Lab work

A - Aerobic/Anaerobic regulation system

Objective : obtaining BBa_K1155000

1 - Electrophoresis of the digestion of BBa_K1155000 by NotI, HpaI, MluI

Sheng, Zhou

PCRPS050813.jpg
  • Well 1, 2 : 5µL of BBa_K1155000 digested by Not I+1µL of 6X loading dye
  • Well 3, 4 : 5µL of BBa_K1155000 digested by Hpa I+1µL of 6X loading dye
  • Well 5 : 6µL of DNA Ladder
  • Well 6, 7 : 5µL of BBa_K1155000 digested by Mlu I+1µL of 6X loading dye
  • Gel 0.8%

Expected sizes :

  • BBa_K1155000 : 111bp

We didn't obtain fragments at the right size. We will do digestions again.

2 - Extraction of BBa_K1155000 from DH5α

Abdou

Transformation of 07/04 works. We will extract DNA plasmid.

Protocol : High copy plamid extraction

Nanodrop :

  • BBa_K1155000 : 761ng/µl

By using NanoDrop, we found some errors which could be due to device malfunctional issue.

Objective : obtaining BBa_K1155003, BBa_K1155007

1 - Extraction of BBa_K592009 and BBa_I732017 from DH5α

Abdou

Transformation of 07/04 works. We will extract DNA plasmid.

Protocol : High copy plamid extraction

Nanodrop :

  • BBa_K592009 : 229ng/µl

By using NanoDrop, we found some errors which could be due to device malfunctional issue.


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