Team:Warsaw/Calendar-Main/19 June 2008

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Preparation of constructs with OmpA protein fusions

Michał K.

Inoculation of separate transformant colonies from plates with ligations: pACYC177+OmpA_alpha pACYC177+OmpA_omega (liquid LB + kanamycin).

Blue/white test

Piotr

  • The analysis of lacZ sequence from pZC
    There is no mutation in beta-galactosidase gene. Why the colonys are white???
  • Colony PCR.
    Template: DNA isolated from white colonies
    Primers: pZCseqL and pZCseqR

    Colony PCR program
    TemperatureTimeNo. of cycles
    94°C4:00
    94°C0:3028 cycles
    48°C0:45
    72°C1:30
    72°C10:00
    4°Cinfinite


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