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Team:Paris Saclay/Notebook/July/23
From 2013.igem.org
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* Well 1 to 3: 30µL of BBa_I732017 digested by EcoRI/SpeI+6µL of 6X loading dye | * Well 1 to 3: 30µL of BBa_I732017 digested by EcoRI/SpeI+6µL of 6X loading dye | ||
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* RBS-LacZ : 3093bp | * RBS-LacZ : 3093bp | ||
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| - | * Term : | + | * pSB3A2 : 2079bp |
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| - | * RBS-LacZ : | + | * RBS-LacZ : 3093bp |
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* Well 1 : 6µL of DNA 6X loading dye | * Well 1 : 6µL of DNA 6X loading dye | ||
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* Gel : 0.8% | * Gel : 0.8% | ||
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Latest revision as of 00:15, 5 October 2013
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
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Expected sizes :
- RBS-LacZ : 3093bp
- Term : 150bp
- pSB3A2 : 2079bp
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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
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Expected sizes :
- RBS-LacZ : 3093bp
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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
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Expected sizes :
- RBS-LacZ : 3093bp
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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
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We can't see anything. The purification wasn't good. We will do cultures again. |
8 - Liquid culture of BBa_I732017 and BBa_K592009 in DH5α
Abdou, Xavier
We let our preparations at 37°C, 150rpm with 5mL of LB and 5mL of chloramphenicol antibiotic.
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