Team:Warsaw/Calendar-Main/24 June 2008

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'''Preparation of pMPMT5-AID+AIDT7 construct'''<br>
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<h3>Preparation of pMPMT5-AID+AIDT7 construct</h3>
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PCR - translation fusion: AID + T7 RNA-polimerase <br>
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<p>PCR - translation fusion: AID + T7 RNA-polimerase <br>
Primers:  
Primers:  
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<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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Template DNA: AID for translation fusion and T7 RNA-polimerase for translation fusion<br>
Template DNA: AID for translation fusion and T7 RNA-polimerase for translation fusion<br>
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Annealing temperature: 55 °C<br>
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Elongation temperature: 55&deg;C<br>
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Annealing time: 4 minutes<br>
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Elongation time: 4 minutes<br>
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Number of cycles: 20 and 25 (2 repeats)<br>
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Number of cycles: 20 and 25 (2 repeats)</p>
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Gel electrophoresis and DNA isolation from proper band (obtained with 20 cycles) ,br
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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>
<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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Revision as of 23:12, 1 October 2008

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Preparation of pMPMT5-AID+AIDT7 construct

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)

Gel electrophoresis and DNA isolation from proper band (obtained with 20 cycles)

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

https://static.igem.org/mediawiki/2008/0/08/Tranlacyjna_pcr_29_05_2008.jpg

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