Team:Warsaw/Calendar-Main/2 June 2008

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<h3>Preparation of pMPMT5-AID+AIDT7 construct<br>
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Michał K.</h3>
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<h4>Michał K.</h4>
<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>
<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>

Revision as of 20:29, 10 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

Blue/white and rifampicin test

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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