Team:Warsaw/Calendar-Main/18 June 2008

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<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>  
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<li> Gel electrophoresis and <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#DNA_isolation_from_agarose_gel">isolation</a> of proper band (3200 bp).</li>
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<li> Gel electrophoresis and <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#DNA_purification_after_enzymatic_reaction">isolation</a> of proper band (3200 bp).</li>
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<li>Clean-up of OmpA_alpha and OmpA_omega digest products. </li>
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<li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#DNA_isolation_from_agarose_gel">Clean-up</a> of OmpA_alpha and OmpA_omega digest products. </li>
<li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#ligation">Ligation</a> of purified DNA (pACYC177 with OmpA_alpha and OmpA_omega DNA fragments).</li>
<li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#ligation">Ligation</a> of purified DNA (pACYC177 with OmpA_alpha and OmpA_omega DNA fragments).</li>
<li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#chemotransform">Transformation</a> of <i>E.coli</i> <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">TOP10</a> with ligation products.</li>
<li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#chemotransform">Transformation</a> of <i>E.coli</i> <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">TOP10</a> with ligation products.</li>

Revision as of 21:13, 9 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.

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