Team:Warsaw/Calendar-Main/22 July 2008

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<h3>Cloning of protein A DNA to OmpA constructs</h3>
 
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<ol><li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#plasmid_DNA_isolation">Isolation of plasmids</a> from cultures inocluated on previous day (pACYC177+OmpA_A_omega).</li>
 
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<li>Control <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest">digest</a> of isolated plasmids with BamHI and SacI. </li></ol>
 
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<h3>Cloning of protein Z DNA to pET15b-OmpA-omega in place of OmpA<br>
 
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Paweł</h3>
 
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<ol><li>Results of ligation: 6 colonies grown.</li>
 
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<li>Each colony cultured overnight in LB + ampicillin</li></ol>
 
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<h3>Cloning omega-A fusion on pKS (second attempt)</h3>
 
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<h4>Michał L., Ewa, Marcin:</h4>
 
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<li>Isolation of plasmids from transformants</li>
 
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<li>Digest of plasmids with SacI and NotI</li>
 
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<li>Gel electrophoresis of products</li>
 
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<li>2 selected clones were send to DNA sequencing lab</li>
 
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<h3>Cloning of protein Z DNA to pET15b-OmpA-omega in place of OmpA<br>
<h3>Cloning of protein Z DNA to pET15b-OmpA-omega in place of OmpA<br>
Paweł</h3>
Paweł</h3>

Revision as of 18:39, 11 October 2008

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Cloning of protein Z DNA to pET15b-OmpA-omega in place of OmpA
Paweł

  1. Isolation of plasmids from cultures inocluated on previous day
  2. Control digest of isolated plasmids with NdeI and NotI. Zero positive results obtained - just empty vectors
  3. Another overnight ligation of pET15b-OmpA-omega (NdeI/NotI cutted) with protein Z DNA


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