Team:Warsaw/Calendar-Main/8 July 2008

From 2008.igem.org

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<li> <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#electrotransform">Transformation</a> of <i>E. coli</i> <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top">TOP10</a>  strain with ligation from previous day </li>
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<li> <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#electrotransform">Transformation</a> of <i>E. coli</i> <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top">TOP10</a>  strain with ligation from previous day. </li>
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<li> Transformants plating on LB + kanamycin</li></ol></p>
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<li> Transformants plating on LB + kanamycin.</li></ol></p>
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<h3>Preparation of construct pKS with A protein<br>
<h3>Preparation of construct pKS with A protein<br>
Michał L., Marcin:</h3>
Michał L., Marcin:</h3>
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<li> Inactivation of digestion enzymes and CIAP</li>
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<li> Inactivation of digestion enzymes and CIAP.</li>
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<li> <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#ligation">Ligation</a> of digested PCR product and pKS 2h at room temperature</li>
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<li> <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#ligation">Ligation</a> of digested PCR product and pKS 2h at room temperature.</li>
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<li> <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#electrotransform">Transformation</a> of <i>E. coli</i> <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top">TOP10</a> strain with 7 µl of ligation mix</li>
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<li> <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#electrotransform">Transformation</a> of <i>E. coli</i> <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top">TOP10</a> strain with 7 µl of ligation mix.</li>
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<li> Transformants plating on LB + ampicillin + X-gal + IPTG</li></ol></p>
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<li> Transformants plating on LB + ampicillin + X-gal + IPTG.</li></ol></p>
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Revision as of 22:35, 1 October 2008

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Preparation of constructs with OmpA protein fusions
Piotr

  1. Transformation of E. coli TOP10 strain with ligation from previous day.
  2. 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 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.