Team:Warsaw/Calendar-Main/21 August 2008

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<h3>Tests for ampicillin resistance given by protein added to medium</h3><h4>Piotr</h4>
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<tr><th>OmpA variant ("hunter")</th><th>"Prey" variant</th><th>Growth with prey</th><th>Growth without prey</th></tr>
<tr><th>OmpA variant ("hunter")</th><th>"Prey" variant</th><th>Growth with prey</th><th>Growth without prey</th></tr>

Revision as of 17:17, 20 October 2008

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Tests for ampicillin resistance given by protein added to medium

Piotr


OmpA variant ("hunter")"Prey" variantGrowth with preyGrowth without prey
OmpA-alphaHis+Z+omega++
OmpA-alphaHis+Z+alfa++
OmpA-A-alphaHis+Z+omega+-
OmpA-Z-omegaHis+A+alpha+-
Ompa-A-omegaHis+Z+alpha+-
OmpA-omegaHis+Z+alpha+-
Ompa-Z-alphaHis+Z+omega+-
OmpA_Adelta_alphaHis+Z+omega+-
OmpA_Adelta_omegaHis+Z+alpha+-
OmpA_omega_AdeltaHis+Z+alpha+-
OmpA-omega-A-alphano++

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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