Team:Warsaw/Calendar-Main/24 June 2008

From 2008.igem.org

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<h3>PCR results <br>Michał K. </h3>
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<p>Sequencing determined that all white colonies have intact lacZ gene.</p>
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<p>Reason of white phenotype unknown.</p>
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<p>We suppose that it is due to chromosomal mutation.</p>
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<p>We need another reporter system (not splitted to chromosomal and plasmid part --> GFP? ).</p>
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<h3>PCR results</h3>
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Still no success. We need to run gradient PCR to find optimal reaction conditions.  
Still no success. We need to run gradient PCR to find optimal reaction conditions.  
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<h3>Preparation of pMPMT5-AID+AIDT7 construct</h3>
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<h3>Preparation of pMPMT5-AID+AIDT7 construct<br>Michał K. </h3>
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<li><a href=<"https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#pcr>PCR</a> - translation fusion: AID + T7 RNA-polymerase <br>
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<li><a href=<"https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#pcr>PCR</a> - translational fusion: AID + T7 RNA-polymerase <br>
Primers:  
Primers:  
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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>
<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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</ol>
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<h3>We need a title here</h3>
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<p>Sequencing determined that all white colonies have intact lacZ gene.</p>
 +
<p>Reason of white phenotype unknown.</p>
 +
<p>We suppose that it is due to chromosomal mutation.</p>
 +
<p>We need another reporter system (not splitted to chromosomal and plasmid part --> GFP? ).</p>
 +

Revision as of 15:29, 9 October 2008

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PCR results
Michał K.

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

Preparation of pMPMT5-AID+AIDT7 construct
Michał K.

  1. PCR - translational fusion: AID + T7 RNA-polymerase
    Primers: AIDlNrH and T7pXbSal
    Template DNA: AID for translation fusion and T7 RNA-polymerase for translation fusion
    Elongation temperature: 55°C
    Elongation time: 4 minutes
    Number of cycles: 20 and 25 (2 repeats)
  2. Gel electrophoresis and DNA isolation from proper band (obtained with 20 cycles)

We need a title here

Sequencing determined that all white colonies 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? ).

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