Team:Warsaw/Calendar-Main/10 July 2008

From 2008.igem.org

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<li> Control <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#digest">digest</a> of isolated plasmids with BamHI and NotI (BamHI buffer).  
<li> Control <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#digest">digest</a> of isolated plasmids with BamHI and NotI (BamHI buffer).  
</li>
</li>
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<li> Gel electrophoresis - we confirmed <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-omega>pACYC177 + OmpA_omega</a>. We didn't obtain <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-alpha>pACYC177 + OmpA_alpha</a> probably because of a mistake in plating. </li>
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<li> Gel electrophoresis - we confirmed <a href="https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-omega">pACYC177 + OmpA_omega</a>. We didn't obtain <a href="https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-alpha">pACYC177 + OmpA_alpha</a> probably because of a mistake in plating. </li>
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<li> <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#ligation">Ligation</a> of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177>pACYC177</a> and OmpA_alpha (1 hr).</li>
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<li> <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#ligation">Ligation</a> of <a href="https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177">pACYC177</a> and OmpA_alpha (1 hr).</li>
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<li> Transformation of <i>E. coli</i> <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">TOP10</a> strain with ligation products: <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-alpha>pACYC177 and OmpA_alpha</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#top10">TOP10</a> strain with ligation products: <a href="https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-alpha">pACYC177 and OmpA_alpha</a>.</li>
<li> Transformants plating on LB + kanamycin.</li>
<li> Transformants plating on LB + kanamycin.</li>
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<h3>Preparation of construct <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pKSII+A>pKS with A protein</a></h3>
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<h3>Preparation of construct <a href="https://2008.igem.org/Wiki/Team:Warsaw/vectors/pKSII+A">pKS with A protein</a></h3>
<h4>Michał L., Marcin</h4>
<h4>Michał L., Marcin</h4>
<p><ol>
<p><ol>

Revision as of 19:32, 17 October 2008

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Preparation of constructs with OmpA protein fusions

Michał K.

  1. Isolation of plasmids from cultures inocluated on previous day.
  2. Control digest of isolated plasmids with BamHI and NotI (BamHI buffer).
  3. Gel electrophoresis - we confirmed pACYC177 + OmpA_omega. We didn't obtain pACYC177 + OmpA_alpha probably because of a mistake in plating.
  4. Ligation of pACYC177 and OmpA_alpha (1 hr).
  5. Transformation of E. coli TOP10 strain with ligation products: pACYC177 and OmpA_alpha.
  6. Transformants plating on LB + kanamycin.

Preparation of construct pKS with A protein

Michał L., Marcin

  1. Inactivation of digestion enzymes and CIAP.
  2. Ligation of digested PCR product and pKS for 2h at room temperature.
  3. Transformation of E. coli TOP10 strain with 7 µl of ligation mix.
  4. Transformants plating on LB + ampicillin + X-gal + IPTG.