Team:Warsaw/Calendar-Main/18 June 2008

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<h3>Preparation of constructs with OmpA protein fusions<br>
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<h3>Preparation of constructs with OmpA protein fusions</h3><h4>
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Michał K.</h3>
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Michał K.</h4>
<ol><li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#plasmid_DNA_isolation">Isolation</a> of pACYC177.</li>
<ol><li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#plasmid_DNA_isolation">Isolation</a> of pACYC177.</li>
<li><a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#digest">Digest</a> of purified pACYC177 with NdeI and BamHI. </li>  
<li><a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#digest">Digest</a> of purified pACYC177 with NdeI and BamHI. </li>  

Revision as of 10:20, 11 October 2008

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Preparation of constructs with OmpA protein fusions

Michał K.

  1. Isolation of pACYC177.
  2. Digest of purified pACYC177 with NdeI and BamHI.
  3. Gel electrophoresis and isolation of proper band (3200 bp).
  4. Clean-up of OmpA_alpha and OmpA_omega digest products.
  5. Ligation of purified DNA (pACYC177 with OmpA_alpha and OmpA_omega DNA fragments).
  6. Transformation of E.coli TOP10 with ligation products.
  7. Plating transformants on LB+tetracycline.

Blue/white and rifampicin test

Michał L., Ewa, Marcin:

Counting of blue colonies. Same results as on 12/06/2008.