Team:Warsaw/Calendar-Main/28 July 2008

From 2008.igem.org

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<p>Sequencing confirms that we have obtained construct with omega-A fusion, but A in our constructs turned out to be trunctated and contain only one from two highly similar domains.</p>
<p>Sequencing confirms that we have obtained construct with omega-A fusion, but A in our constructs turned out to be trunctated and contain only one from two highly similar domains.</p>
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<h3>Cloning of omega_A DNA fragment to pACYC177+OmpA_alpha</h3><h4>Michał K.</h4>
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<ol><li>DNA fragment of omega_A isolated from gel on 20 July <a href=https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest>digestion</a> with SacI and NotI.</li>
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<li><a href=https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest>clean-up</a>Clean-up of digested DNA fragment.</li>
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<li><a href=https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest>Digestion</a> (SacI and NotI) and dephosphorylation (CIAP) of pACYC177+OmpA_alpha. </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">gel-out</a> of 4300 bp band. </li>
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<li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#ligation">Ligation</a> of DNA fragments from 2. and 4. (1 hr)</li>
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<li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#chemotransform">Transformation</a> of E. coli <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">TOP10</a> strain with ligation.</li>
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<li>Transformants plating on LB + kanamycin.</li></ol>
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</p>
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<h3>Checking the expression of omp_omega_A_alpha and omp_A_alpha<br>
<h3>Checking the expression of omp_omega_A_alpha and omp_A_alpha<br>

Revision as of 21:16, 11 October 2008

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Cloning omega-A fusion on pKS (second attempt)

Michał L., Ewa, Marcin

Sequencing confirms that we have obtained construct with omega-A fusion, but A in our constructs turned out to be trunctated and contain only one from two highly similar domains.

Checking the expression of omp_omega_A_alpha and omp_A_alpha
Piotr

  1. Inoculation of omp_omega_A_alpha and omp_A_alpha with inductor (0,5 mmol/mL IPTG) and negative control without IPTG.

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