Team:Warsaw/Calendar-Main/27 August 2008

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<h3>Cloning of protein A DNA to pET15b+Omp-alfa plasmid in place of OmpA<br>Antoni</h3>
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<h3>Cloning of protein A DNA to pET15b+OmpA-alfa plasmid in place of OmpA<br>Antoni</h3>
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<li> <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#digest">Digest</a> pET15b+OmpA_alpha and pGeneart+A with NdeI and NotI, pET15b+OmpA_alpha<a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#removing"> dephosphorylation with CIAP</a> </li> </li>
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<li> Colony PCR on colonies from plates with transformations pGeneart+A Primers used:
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<li> Gel electrophoresis of digested plasmids and <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#DNA_isolation_from_agarose_gel">gel-out</a> of proper bands  </li>
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<li> <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#ligation">Ligation</a> of pET15b+alfa and A </li>
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<li> Transformation of <i>E. coli</i> <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">TOP10</a>. </li>
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<li> Transformants plating on LB + ampicillin.</li>
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<a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#AP+NotI">AP+NotI</a> and <a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#AL+SacI">AL+SacI</a>
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<li> Confirmed transformant colonies inoculated to liquid LB with ampicillin
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Revision as of 15:11, 9 October 2008

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Cloning of protein A DNA to pET15b+OmpA-alfa plasmid in place of OmpA
Antoni

  1. Colony PCR on colonies from plates with transformations pGeneart+A Primers used: AP+NotI and AL+SacI
  2. Confirmed transformant colonies inoculated to liquid LB with ampicillin



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