Team:Warsaw/Calendar-Main/27 August 2008

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<h3>Cloning of protein A DNA to pET15b+Omp-alfa plasmid<br>Antoni</h3>
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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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<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>
<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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Cloning of protein A DNA to pET15b+Omp-alfa plasmid in place of OmpA
Antoni

  1. Digest pET15b+OmpA_alpha and pGeneart+A with NdeI and NotI, pET15b+OmpA_alpha dephosphorylation with CIAP
  2. Gel electrophoresis of digested plasmids and gel-out of proper bands
  3. Ligation of pET15b+alfa and A
  4. Transformation of E. coli TOP10.
  5. Transformants plating on LB + ampicillin.

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