Team:Warsaw/Calendar-Main/24 July 2008

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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>
<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>
<li> <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#ligation">Ligation</a> of pET15b+alfa and A </li>
<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#top">TOP10</a>. </li>
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<li> Transformants plating on LB + ampitilin.</li>
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Revision as of 20:22, 2 October 2008

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1. Separate transformant colonies (tranformation from previous day) inoculated to liquid LB with kanamycin.

Antoni:

1. Preparation of chemocompetent bacteria E. coli TOP10.

Cloning of protein A DNA to pET15b+Omp-alfa plasmid
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 + ampitilin.

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