Team:Warsaw/Calendar-Main/24 June 2008

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<h3>PCR results<br>
 
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Michał L., Ewa, Marcin</h3>
 
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Still no success. We need to run gradient PCR to find optimal reaction conditions.  
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<h3>Preparation of alpha-A and omega-A fusions</h3>
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<h4>Michał L., Ewa, Marcin</h4>
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<p>Still no success. We need to run gradient PCR to find optimal reaction conditions.</p>
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<h3>Preparation of constructs with OmpA protein fusions<br>
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Michał K.</h3>
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<h3>Preparation of constructs: OmpA_alpha and OmpA_omega</h3>
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<h4>Michał K.</h4>
<ol><li> PCR on isolated plasmids with <a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#OmpaL_N">OmpaL_N</a> and <a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#OmpaP_link">OmpaP_link</a>
<ol><li> PCR on isolated plasmids with <a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#OmpaL_N">OmpaL_N</a> and <a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#OmpaP_link">OmpaP_link</a>
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  primers</li>
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  primers.</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">DNA isolation</a> from proper band (obtained with 20 cycles)</li>
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<li>Gel electrophoresis (<a href="https://2008.igem.org/wiki/index.php?title=Team:Warsaw/Calendar-Main/24_June_2008#fig1">Fig. 1</a>, no proper clones found).</li>
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<h3> Blue/white and rifampicin test </h3>
 
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<p>Sequencing determined that all white colonies from <a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/16_June_2008">16 June</a> have intact lacZ gene. Reason of white phenotype unknown. We suppose that it is due to chromosomal mutation. We need another reporter system (not splitted to chromosomal and plasmid part --> GFP? ).</p>
 
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<a name="fig1"><img src="https://static.igem.org/mediawiki/2008/7/78/Omp_colony_PCR_WAW.jpg" width=350/></a>
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<var><b>Fig. 1.</b> PCR product - searching for fusions with OmpA protein. Lower lanes - OmpA_alpha, upper - OmpA_omega.</var>

Latest revision as of 16:53, 28 October 2008

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Preparation of alpha-A and omega-A fusions

Michał L., Ewa, Marcin

Still no success. We need to run gradient PCR to find optimal reaction conditions.

Preparation of constructs: OmpA_alpha and OmpA_omega

Michał K.

  1. PCR on isolated plasmids with OmpaL_N and OmpaP_link primers.
  2. Gel electrophoresis (Fig. 1, no proper clones found).

Fig. 1. PCR product - searching for fusions with OmpA protein. Lower lanes - OmpA_alpha, upper - OmpA_omega.

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