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Team:Paris Saclay/Notebook/July/3
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(→1 - Colony PCR of Bba_K1155000 to check good insertion of Pfnr in PSB1C3) |
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==='''A - Aerobic/Anaerobic regulation system'''=== | ==='''A - Aerobic/Anaerobic regulation system'''=== | ||
| - | ===='''Objective : obtaining | + | ===='''Objective : obtaining BBa_K1155000'''==== |
| - | ===='''1 - Colony PCR of | + | ===='''1 - Colony PCR of BBa_K1155000 to check good insertion of Pndh* in PSB1C3'''==== |
Abdou, Sheng, Zhou | Abdou, Sheng, Zhou | ||
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| - | Colonies count for | + | Colonies count for BBa_K1155000 : |
* Standard concentration : | * Standard concentration : | ||
| - | ** | + | ** BBa_K1155000 : 0 |
* High concentration : | * High concentration : | ||
| - | ** | + | ** BBa_K1155000 : 2 |
[[File:Ps0307jour.jpg|300px]] | [[File:Ps0307jour.jpg|300px]] | ||
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[[File:PSPCR0307.jpg|400px]] | [[File:PSPCR0307.jpg|400px]] | ||
| - | ===='''2 - Culture of | + | ===='''2 - Culture of BBa_K1155000 '''==== |
Sheng | Sheng | ||
| - | We made new cultures by streaking 2 colonies of 07/02/13 culture of | + | We made new cultures by streaking 2 colonies of 07/02/13 culture of BBa_K1155000. We also did liquid culture of each one. |
We let culture at 37°C. | We let culture at 37°C. | ||
| - | ===='''Objective : obtaining | + | ===='''Objective : obtaining BBa_K1155003, BBa_K1155007'''==== |
| - | ===='''1 - Culture of | + | ===='''1 - Culture of BBa_I732017, BBa_K592009 '''==== |
Sheng | Sheng | ||
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|} | |} | ||
| - | We made new cultures by streaking 2 colonies of 07/02/13 culture of | + | We made new cultures by streaking 2 colonies of 07/02/13 culture of BBa_I732017, BBa_K592009. We also did liquid culture of each one. |
We let culture at 37°C. | We let culture at 37°C. | ||
Revision as of 10:05, 30 September 2013
Contents |
Notebook : July 3
Lab work
A - Aerobic/Anaerobic regulation system
Objective : obtaining BBa_K1155000
1 - Colony PCR of BBa_K1155000 to check good insertion of Pndh* in PSB1C3
Abdou, Sheng, Zhou
|
Tranformation of 07/02/13 works. We will do a Colony PCR of colonies. |
Colonies count for BBa_K1155000 :
- Standard concentration :
- BBa_K1155000 : 0
- High concentration :
- BBa_K1155000 : 2
Primer and PCR :
VF2, VR, Pfnr_up, Pfnr_down are four oligos that we used for plasmid amplification. We used tree combinaisons VF/VR, VF/Pfnr_Down, Pfnr_Up/VR. If the promoter Pfnr insert PSB1C3 plasmid successfully, tree fragments with specific size will be amplified.
We mix each colony with 20µL of H2O.
Used quantities :
- DNA : 2µL
- Mix : (it was divided in tubes for 4 different colonies for each oligo combinaison with 23µL of mix in each tube)
- VF or Pfnr_Up : 6µL
- VR or Pfnr_Down or VR : 6µL
- dNTP : 6µL
- Buffer Dream Taq : 30µL
- Dream Taq : 6µL
- H2O : 246µL
2 - Culture of BBa_K1155000
Sheng
We made new cultures by streaking 2 colonies of 07/02/13 culture of BBa_K1155000. We also did liquid culture of each one. We let culture at 37°C.
Objective : obtaining BBa_K1155003, BBa_K1155007
1 - Culture of BBa_I732017, BBa_K592009
Sheng
|
Tranformation of 07/02/13 works. We will do new cultures. |
We made new cultures by streaking 2 colonies of 07/02/13 culture of BBa_I732017, BBa_K592009. We also did liquid culture of each one. We let culture at 37°C.
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