Team:Paris Saclay/Notebook/July/8

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Notebook : July 8

Lab work

A - Aerobic/Anaerobic regulation system

Objective : Bba_K1155003, Bba_K1155007

1 - Digestion of Bba_K592009, Bba_I732017 by NotI, XhoI, EcoRI, EcoRI/PstI

Abou, Anaïs, Sheng, Zhou

Used quantities :

  • NotI :
    • Bba_K592009 : 2µL
    • Buffer orange : 2µL
    • NotI : 0.5µL
    • H2O : 15.5µL
  • XhoI :
    • Bba_K592009 : 2µL
    • Buffer red : 2µL
    • XhoI : 0.5µL
    • H2O : 15.5µL
  • EcoRI :
    • Bba_K592009 : 2µL
    • Buffer orange : 2µL
    • EcoRI : 0.5µL
    • H2O : 15.5µL
  • EcoRI/PstI :
    • Bba_I732017 : 2µL
    • Buffer orange : 2µL
    • EcoRI : 0.5µL
    • PstI : 0.5µL
    • H2O : 15µL

We let our digestion 1h30 at 37°C.

2 - Electrophoresis of the digestion of Bba_K592009, Bba_I732017 by NotI, XhoI, EcoRI, EcoRI/PstI

Abdou, Anaïs, Sheng, Zhou

[[]]
  • Well 1 : 6µL DNA Ladder
  • Well 2 : 5µL Bba_K592009 clone 1 digested by EcoRI+1µl of 6X loading dye
  • Well 3 : 5µL Bba_K592009 clone 2 digested by EcoRI+1µl of 6X loading dye
  • Well 4 : 5µL Bba_K592009 clone 1 digested by NotI+1µl of 6X loading dye
  • Well 5 : 5µL Bba_K592009 clone 2 digested by NotI+1µl of 6X loading dye
  • Well 6 : 5µL Bba_K592009 clone 1 digested by XhoI+1µl of 6X loading dye
  • Well 7 : 5µL Bba_K592009 clone 2 digested by XhoI+1µl of 6X loading dye
  • Well 8 : 5µL Bba_I732017 clone 1 digested by EcoRI/PstI+1µl of 6X loading dye
  • Well 9 : 5µL Bba_I732017 clone 2 digested by EcoRI/PstI+1µl of 6X loading dye
  • Well 10 : -
  • Well 11 : -
  • Well 12 : 6µL DNA Ladder
  • Gel : 1%

Expected sizes :

  • Bba_K592009 digested by NotI : 2046bp + 693bp
  • Bba_K592009 digested by XhoI : 1842bp + 892bp

we obtain fragments at the right size. Our extraction was good.

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