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Team:Paris Saclay/Notebook/July/16
From 2013.igem.org
Notebook : July 16
Lab work
A - Aerobic/Anaerobic regulation system
Objective : obtaining Bba_K1155003, Bba_K1155007
1 - Extraction of Bba_B0015, Bba_B0017, Bba_I732019 from DH5α
Zhou
Protocol : High copy plamid extraction
2 - Digestion of Bba_B0015, Bba_B0017, Bba_I732019 by EcoRI/PstI
Anaïs
- DNA : 5µL
- Buffer FD : 2µL
- EcoRI FD : 1µL
- PstI FD : 1µL
- H2O : 21µL
We let our digestion 1h30 at 37°C.
Objective : obtaining biobricks in PSB3K3
1 - Digestion of PSB3K3 by EcoRI/PstI
Anaïs
Used quantities :
- DNA : 5µL
- Buffer FD : 2µL
- EcoRI FD : 1µL
- PstI FD : 1µL
- H2O : 11µL
We let our digestion 1h30 at 37°C.
2 - Electrophoresis of the digestion of PSB3K3 by EcoRI/PstI
Sheng
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Expected sizes :
- PSB3K3 : 2750bp
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We obtain fragments at the right size. |
3 - Gel purification of electrophoresis of the digestion of PSB3K3 by EcoRI/PstI
Abdou
Protocol : [http://www.mn-net.com/tabid/1452/default.aspx Gel purification ]
B - PCB sensor system
Objective : obtaining Bba_K1155001, Bba_K1155002, BphR2 in PSB1C3
1 - Electrophoresis of the digestion of Bba_K1155001, Bba_K1155002, BphR2 in PSB1C3
Anaïs, Zhou
- Bba_K1155001 :
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Expected size :
- Bba_K1155001 digested by EcoRI/PstI : 2037bp + 333bp
- Bba_K1155001 digested by SacII : 2370bp
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In the first well, we obtain fragment at the right size. We will amplify Bba_K1155001. |
- Bba_K1155002 :
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Expected size :
- Bba_K1155002 digested by EcoRI/PstI : ...
- Bba_K1155002 digested by SacII : ...
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We didn't obtain fragments at the right size. We will do the ligation again. |
- BphR2 in PSB1C3 :
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Expected size :
- BphR2 digested by EcoRI/PstI : ...
- BphR2 digested by SacII : ...
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We didn't obtain fragments at the right size. After reading the sequence again, we find an digestion site in the middle of our biobrick. We will do a Gibson assembly to modify this site. |
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