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Team:Paris Saclay/Notebook/July/16
From 2013.igem.org
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| - | * Well 1 to 8 : 2µL | + | * Well 1 to 8 : 2µL BphR2 in PSB1C3 digested by EcoRI/PstI+2µl of 6X loading dye |
* Well 9 : 6µL DNA Ladder | * Well 9 : 6µL DNA Ladder | ||
| - | * Well 10 to 17 : 2µL | + | * Well 10 to 17 : 2µL BphR2 in PSB1C3 digested by SacII+2µl of 6X loading dye |
* Gel : 1.5% | * Gel : 1.5% | ||
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Expected size : | Expected size : | ||
| - | * | + | * BphR2 digested by EcoRI/PstI : ... |
| - | * | + | * BphR2 digested by SacII : ... |
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| - | We didn't obtain fragments at the right size. We will do | + | 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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|[[Team:Paris_Saclay/Notebook|<big>Back to calendar</big>]] | |[[Team:Paris_Saclay/Notebook|<big>Back to calendar</big>]] | ||
| - | |[[Team:Paris Saclay/Notebook/July/17|<big> | + | |[[Team:Paris Saclay/Notebook/July/17|<big>Next day</big>]] |
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{{Team:Paris_Saclay/incl_fin}} | {{Team:Paris_Saclay/incl_fin}} | ||
Revision as of 19:15, 26 September 2013
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
| [[]] |
|
Expected sizes :
- PSB3K3 : 2750bp
|
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 :
| [[]] |
|
Expected size :
- Bba_K1155001 digested by EcoRI/PstI : 2037bp + 333bp
- Bba_K1155001 digested by SacII : 2370bp
|
In the first well, we obtain fragment at the right size. We will amplify Bba_K1155001. |
- Bba_K1155002 :
| [[]] |
|
Expected size :
- Bba_K1155002 digested by EcoRI/PstI : ...
- Bba_K1155002 digested by SacII : ...
|
We didn't obtain fragments at the right size. We will do the ligation again. |
- BphR2 in PSB1C3 :
| [[]] |
|
Expected size :
- BphR2 digested by EcoRI/PstI : ...
- BphR2 digested by SacII : ...
|
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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