Team:Paris Saclay/Notebook/July/3

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===='''2 - Colony PCR of Bba_K1155000, Bba_I732017, Bba_K592009'''====
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===='''2 - Culture of Bba_K1155000, Bba_I732017, Bba_K592009 '''====
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Sheng
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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. 
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We let culture at 37°C.
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Revision as of 14:39, 22 September 2013

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

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


PSprimer07.jpg


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

PsPCR2908.jpg

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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