Team:Warsaw/Calendar-Main/26 August 2008

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<h3>Cloning of protein A DNA to pET15b+OmpA-alfa plasmid in place of OmpA<br>Antoni</h3>
<h3>Cloning of protein A DNA to pET15b+OmpA-alfa plasmid in place of OmpA<br>Antoni</h3>
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<li> Colony PCR on colonies from plates with transformations pGeneart+A Primers used:
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<li> Colony <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#pcr">PCR</a> on colonies from plates with transformations pGeneart+A Primers used:
<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></li>
<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></li>

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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. Lack of confirmed transformant colonies

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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