Team:Warsaw/Calendar-Main/2 June 2008

From 2008.igem.org

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<h4>Michał L., Ewa, Marcin:</h4>  
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<h3>Preparation of pMPMT5-AID+AIDT7 construct<br>
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Michał K.</h3>
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<p><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#pcr">PCR</a> for AID transcription fusion with 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#chemotransform">Transformation</a> of <i>E. coli</i> <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">TOP10</a>  with <a href="https://2008.igem.org/Wiki/Team:Warsaw/vectors/pZC320">pZC320</a>.</li>
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<li> Plating transformants on LB + ampicillin(30μg/mL) + X-gal + IPTG.</li></ol><br></p>
 
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<h3>Preparation of pMPMT5-AID+AIDT7 construct</h3>
 
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<h4>Michał K.:</h4>
 
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<ol><li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#pcr">PCR</a> for AID transcription fusion with translation fusion AID+T7 RNA-polymerase. <br></li>
 
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<p>
 
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Primers: <br>
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Primers:  
<a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#AIDlNrH">AIDlNrH</a> and <a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#T7pXbSal">T7pXbSal</a><br>
<a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#AIDlNrH">AIDlNrH</a> and <a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#T7pXbSal">T7pXbSal</a><br>
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Number of cycles: 20 and 25
Number of cycles: 20 and 25
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</li></ol>
 
</p>
</p>
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<h4>Michał L., Ewa, Marcin:</h4>
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<p>
 +
<ol>
 +
<li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#chemotransform">Transformation</a> of <i>E. coli</i> <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">TOP10</a>  with <a href="https://2008.igem.org/Wiki/Team:Warsaw/vectors/pZC320">pZC320</a>.</li>
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<li> Plating transformants on LB + ampicillin (30 μg/mL) + X-gal + IPTG.</li></ol><br></p>

Revision as of 14:47, 9 October 2008

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Preparation of pMPMT5-AID+AIDT7 construct
Michał K.

PCR for AID transcription fusion with translation fusion AID+T7 RNA-polymerase.
Primers: AIDlNrH and T7pXbSal
Template DNA: translation fusion AID+T7 RNA-polymerase (pMPM-T5)
Elongation temperature: 55°C
Time: 4 minutes
Number of cycles: 20 and 25

Michał L., Ewa, Marcin:

  1. Transformation of E. coli TOP10 with pZC320.
  2. Plating transformants on LB + ampicillin (30 μg/mL) + X-gal + IPTG.

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