Team:Paris Saclay/Notebook/August/5

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Notebook : August 5

Lab work

A - Aerobic/Anaerobic regulation system

Objective : obtaining Bba_K1155003

1 - Digestion of Bba_K1155003 by EcoRI/PstI

Nadia, XiaoJing

Used quantities :

  • Bba_K1155003 : 5µL
  • EcoRI FD : 1µL
  • PstI FD : 1µL
  • Buffer FD : 3µL
  • H2O : 20µL

We let the digestion at 37°C during 10 minutes ??????

2 - Electrophoresis of the digestion of Bba_K1155003

Damir

File:Ps.jpg|350px]]
  • Well 1 :
  • Well 2 : 6µL of DNA Ladder
  • Well 3 : 30µL of Bba_K1155003 from clone 9 digested by EcoRI/PstI+6µl of 6X loading dye
  • Well 4 : 5µL of Bba_K1155003 from clone 9+1µl of 6X loading dye
  • Well 5 : 30µL of Bba_K1155003 from clone 11 digested by EcoRI/PstI+6µl of 6X loading dye
  • Well 6 : 5µL of Bba_K1155003 from clone 11+1µl of 6X loading dye
  • Well 7 : 30µL of Bba_K1155003 from clone 12 digested by EcoRI/PstI+6µl of 6X loading dye
  • Well 8 : 5µL of Bba_K1155003 from clone 12+1µl of 6X loading dye
  • Gel : 1%

Objective : obtaining Bba_K1155004, Bba_K1155005, Bba_K1155006

1 - Colony PCR of Bba_K1155004, Bba_K115005, Bba_K1155006 in DH5α

Damir, Nadia, XiaoJing

COLONIES PIQUEES DANS 10µL d'eau par Tube !!!!!!!!!!!!!

Used quantities :

  • DNA : 2µL
  • Mix  : (it was divided in 25 tubes for each promotor with 23µL of mix in each on)
    • Oligo 44 : 3.5µL
    • Oligo 45 : 3.5µL
    • Buffer Dream Taq : 70µL
    • dNTP : 28µL
    • Dream Taq : 5µL
    • H2O : 590µL

PCR Program :

PsPCR0508.jpg


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