Team:Warsaw/Calendar-Main/5 July 2008

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<h3>Preparation of constructs with OmpA protein fusions<br>Paweł</h3>
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<h3>Preparation of constructs with OmpA protein fusions<br>
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Piotr:</h3>
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<li> <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#plasmid_DNA_isolation">Isolation</a> of plasmids from bacterial cultures inoculated on previous day (pET15b+OmpA_alpha and pET15b+OmpA_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 plasmids with NotI and BamHI. </li>
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<li> Gel electrophoresis. </li>
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<li> <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#digest">Digest</a> of confirmed clones of pET15b+OmpA_alpha and pET15b+OmpA_omega with NdeI and BamHI. </li>
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<li> <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#digest">Digest</a> of pACYC177 plasmid with NdeI and BamHI, <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#removing">dephosphorylation with CIAP</a>. </li>
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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>Electrophoresis of gel-out products.</li>
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<li> Overnight <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#ligation">ligation</a> of pACYC177 and OmpA_alpha. </li>
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<li> Overnight <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#ligation">ligation</a> of pACYC177 and OmpA_omega. </li>
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Revision as of 21:45, 9 October 2008

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Preparation of constructs with OmpA protein fusions
Piotr:

  1. Digest of confirmed clones of pET15b+OmpA_alpha and pET15b+OmpA_omega with NdeI and BamHI.
  2. Digest of pACYC177 plasmid with NdeI and BamHI, dephosphorylation with CIAP.
  3. Gel electrophoresis of digested plasmids and gel-out of proper bands.
  4. Electrophoresis of gel-out products.
  5. Overnight ligation of pACYC177 and OmpA_alpha.
  6. Overnight ligation of pACYC177 and OmpA_omega.


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