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Team:Paris Saclay/Notebook/July/3
From 2013.igem.org
(→1 - Colony PCR of BBa_K1155000 to check good insertion of Pndh* in pSB1C3) |
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| - | '''[Primers] and PCR :''' | + | '''[https://2013.igem.org/Team:Paris_Saclay/Primers Primers] and PCR :''' |
<p>'''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.''' | <p>'''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.''' </p> | '''If the promoter Pfnr insert PSB1C3 plasmid successfully, tree fragments with specific size will be amplified.''' </p> | ||
Revision as of 17:04, 4 October 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
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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
Primers 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
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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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