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Team:Paris Saclay/Notebook/July/3
From 2013.igem.org
Culture confirmation
We performed another colonies seeding. 2 colonies selected from each Petri dish were seeded in liquid medium with their corresponding antibiotic at 37°C under stirring during 1 night.
Contents |
Notebook : July 1
Lab work
A - Aerobic/Anaerobic regulation system
Objective : obtaining Bba_K1155000
1 - Colony PCR of Bba_K1155000, Bba_I732017, Bba_K592009
Abdou, Sheng, Zhou
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Tranformation of 07/02/13 works. We will do a Colony PCR of colonies. |
Colonies count :
- 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.
COLONIES PIQUEES DANS 20µL d'eau pour chaque colonie.
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, Bba_I732017, Bba_K592009
Sheng
We made we culture by streaking 2 colonies of 07/02/13 culture of Bba_K1155000, Bba_I732017, Bba_K592009. We also did liquid culture of each one. We let culture at 37°C.
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