Team:Warsaw/Calendar-Main/27 September 2008

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<h3>Protein A mutagenesis<br>Paweł</h3>
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<h3>Mutagenesis of protein A</h3><h4>Paweł</h4>
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<p><ol><li>Isolation of plasmid DNA from overnight cultures (it was really tough work...)</li>
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<p><ol><li>Isolation of plasmid DNA from overnight cultures (it was really tough work...).</li>
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<li>Control digestion of isolated plasmids with KpnI and BamHI (if mutagenesis really occured, ~550bp band should be visible)</li>
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<li>Control digestion of isolated plasmids with KpnI and BamHI (if mutagenesis really occured, ~550bp band should be visible).</li>
<li>Results of digestion:</li>
<li>Results of digestion:</li>
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<li>Plasmids which gave positive digestion results prepared for sequencing (one sample from each variant which gave positive result)</li>
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<li>Plasmids which gave positive digestion results prepared for sequencing (one sample from each variant which gave positive result).</li>
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Revision as of 20:14, 13 October 2008

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Mutagenesis of protein A

Paweł

  1. Isolation of plasmid DNA from overnight cultures (it was really tough work...).
  2. Control digestion of isolated plasmids with KpnI and BamHI (if mutagenesis really occured, ~550bp band should be visible).
  3. Results of digestion:
    • ADelL+KpnI and ADelP primers mutagenesis:
      • pACYClac+ompA-A-omega: 0 positive, 5 negative
      • pACYClac+ompa-A-alpha: 3 positive, 7 negative
      • pACYClac+ ompa-alpha-A: 1 positive, 2 negative
    • AMutL+KpnI and AMutP mutagenesis:
      • pACYClac+ompA-A-omega: 4 positive, 16 negative
      • pACYClac+ompa-A-alpha: 4 positive, 8 negative
      • pACYClac+ ompa-alpha-A: 2 positive, 15 negative
  4. Plasmids which gave positive digestion results prepared for sequencing (one sample from each variant which gave positive result).

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