Team:Warsaw/Calendar-Main/24 June 2008

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<h3>Preparation of pMPMT5-AID+AIDT7 construct</h3>
<h3>Preparation of pMPMT5-AID+AIDT7 construct</h3>
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<p>PCR - translation fusion: AID + T7 RNA-polimerase <br>
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<ol>
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<li>PCR - translation fusion: AID + T7 RNA-polimerase <br>
Primers:  
Primers:  
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Elongation time: 4 minutes<br>
Elongation time: 4 minutes<br>
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Number of cycles: 20 and 25 (2 repeats)</p>
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Number of cycles: 20 and 25 (2 repeats)</li>
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<p>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)</p>
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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>
<h3>PCR results of translational fusion for the pMPMT5-AID+AIDT7 construct.<br> Lane 1 - 25 cycles, lane 2 - 20 cycles.</h3>
<h3>PCR results of translational fusion for the pMPMT5-AID+AIDT7 construct.<br> Lane 1 - 25 cycles, lane 2 - 20 cycles.</h3>

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Preparation of pMPMT5-AID+AIDT7 construct

  1. PCR - translation fusion: AID + T7 RNA-polimerase
    Primers: AIDlNrH and T7pXbSal
    Template DNA: AID for translation fusion and T7 RNA-polimerase 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)

    3. PCR results of translational fusion for the pMPMT5-AID+AIDT7 construct.
    Lane 1 - 25 cycles, lane 2 - 20 cycles.

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