Team:Warsaw/Calendar-Main/21 August 2008

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OmpA_A_alpha, OmpA_Adelta_alpha, OmpA_Adelta_omega, OmpA_A_omega, OmpA_omega_Adelta, OmpA_omeg
OmpA variant ("hunter")"Prey" variantGrowth with preyGrowth without prey
OmpA-alphaHis+Z+omeganono
OmpA-alphaHis+Z+alfanono
OmpA-A-alphaHis+Z+omegayesno
OmpA-Z-omegaHis+A+alphayesno
OmpA-omega-A-alphaHis+Z+omegayesno
Ompa-A-omegaHis+Z+alphayesno
Ompa-A-omegaHis+Z+alphayesno
OmpA-omegaHis+A+alphanono
Ompa-Z-alphaHis+A+omegayesno
Ompa-Z-alphaHis+A+omegayesno

Cloning of protein A DNA to GeneArt plasmid

Antoni

  1. Colony PCR on colonies from plates with transformations pGeneart+A.
    Primers used: AP+NotI and AL+SacI.
  2. Gel electrophoresis.
  3. Confirmed transformant colonies inoculated to liquid LB with ampicillin.
  4. Inoculation to liquid LB with ampicillin: pET15b+OmpA-alpha.

Cloning of alpha to pACYC177+OmpA_A_omega and pACYC177+OmpA_Z_omega

Michał K.

  1. Isolation of plasmids from cultures inocluated on previous day.
  2. Control digest of isolated plasmids with FastBamHI and FastAcc65I (Fast Digest buffer).
  3. Gel electrophoresis - no proper clones founded.

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