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Team:Paris Saclay/Notebook/July/31
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(Difference between revisions)
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==='''A - Aerobic/Anaerobic regulation system'''=== | ==='''A - Aerobic/Anaerobic regulation system'''=== | ||
| - | ===='''Objective : obtaining | + | ===='''Objective : obtaining BBa_K1155004, BBa_K1155005, BBa_K155006'''==== |
===='''1 -Digestion of PSB1C3 by EcoRI/PstI'''==== | ===='''1 -Digestion of PSB1C3 by EcoRI/PstI'''==== | ||
| Line 65: | Line 65: | ||
|} | |} | ||
| - | ===='''5 -Transformation of | + | ===='''5 -Transformation of BBa_K1155004, BBa_K1155005, BBa_K1155006'''==== |
Anaïs | Anaïs | ||
| Line 71: | Line 71: | ||
Protocol : [[Team:Paris_Saclay/Protocols/Bacterial transformation|Bacterial transformation]] | Protocol : [[Team:Paris_Saclay/Protocols/Bacterial transformation|Bacterial transformation]] | ||
| - | ===='''Objective : obtaining | + | ===='''Objective : obtaining BBa_K1155007'''==== |
| - | ===='''1 - Digestion of | + | ===='''1 - Digestion of BBa_I732017 by EcoRI/SpeI'''==== |
XiaoJing | XiaoJing | ||
Used quantities : | Used quantities : | ||
| - | * | + | * BBa_I732017 : 41µL |
* Buffer FD : 5µL | * Buffer FD : 5µL | ||
* EcoRI : 2µL | * EcoRI : 2µL | ||
| Line 85: | Line 85: | ||
We let the digestion at 1h30 at 37°C. | We let the digestion at 1h30 at 37°C. | ||
| - | ===='''2 -Electrophoresis to check the digestion of | + | ===='''2 -Electrophoresis to check the digestion of BBa_I732017 by EcoRI/SpeI'''==== |
XiaoJing | XiaoJing | ||
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| style="width:350px;border:1px solid black;vertical-align:top;" | | | style="width:350px;border:1px solid black;vertical-align:top;" | | ||
* Well 1 : 6µL of DNA Ladder | * Well 1 : 6µL of DNA Ladder | ||
| - | * Well 2 et 3 : 40µL of | + | * Well 2 et 3 : 40µL of BBa_I732017 digested by EcoRI/SpeI+8µL 6X loading dye |
* Gel : 0.8% | * Gel : 0.8% | ||
|} | |} | ||
Revision as of 09:32, 30 September 2013
Notebook : July 31
Lab work
A - Aerobic/Anaerobic regulation system
Objective : obtaining BBa_K1155004, BBa_K1155005, BBa_K155006
1 -Digestion of PSB1C3 by EcoRI/PstI
Abdou
Used quantities :
- PSB1C3 : 16µL
- Buffer FD : 2µL
- EcoRI FD : 1µL
- PstI FD : 1µL
We let our digestion 1h30 at 37°C.
2 -Denaturation of EcoRI/PstI used for the digestion of PSB1C3
Abdou
Protocol : Ethanol precipitation
3 -Ligation of NarK, NarG, NirB in PSB1C3
XiaoJing
Used quantities :
- Buffer : 2µL
- NarK, NarG, NirB : 1µL
- PSB1C3 : 2µL
- Ligase : 1µL
- H2O : 14µL
4 -Electrophoresis to check the ligation of NarK, NarG, NirB in PSB1C3
XiaoJing
| [[]] |
|
Expected sizes :
- NarK : ...
- Nar G : ...
- Nir B : ...
|
We obtain fragments at the right size. We will transform them in DH5α. |
5 -Transformation of BBa_K1155004, BBa_K1155005, BBa_K1155006
Anaïs
Protocol : Bacterial transformation
Objective : obtaining BBa_K1155007
1 - Digestion of BBa_I732017 by EcoRI/SpeI
XiaoJing
Used quantities :
- BBa_I732017 : 41µL
- Buffer FD : 5µL
- EcoRI : 2µL
- SpeI : 2µL
We let the digestion at 1h30 at 37°C.
2 -Electrophoresis to check the digestion of BBa_I732017 by EcoRI/SpeI
XiaoJing
| [[]] |
|
Expected sizes :
- RBS-LacZ : 3093 bp
- PSB1A2 : 2079 bp
|
We obtain fragments at the right size. We will purify it. |
3 - Gel purification of digestion of RBS-LacZ
Xavier
Protocol : Gel purification
|
We lost our fragments. We will do the digestion again. |
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