Team:Warsaw/Calendar-Main/26 June 2008

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<h3>Preparation of pMPMT5-AID+AIDT7 construct <br>Michał K.</h3>
<h3>Preparation of pMPMT5-AID+AIDT7 construct <br>Michał K.</h3>
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<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>  strain with <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#ligation">ligation</a> from previous day </li>
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<li>Plating transformants on LB + tetracycline</li></ol>
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<li> Repetition of PCRs, gel-out isolations and digests from <a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/17_June_2008">17 June.</a></li></ol>
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<h3> Polymerase Chain Ligation (PCL) <br>Michał L., Ewa, Marcin</h3>
<h3> Polymerase Chain Ligation (PCL) <br>Michał L., Ewa, Marcin</h3>

Revision as of 21:02, 9 October 2008

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Preparation of pMPMT5-AID+AIDT7 construct
Michał K.

  1. Repetition of PCRs, gel-out isolations and digests from 17 June.

Polymerase Chain Ligation (PCL)
Michał L., Ewa, Marcin

ProductTemplatesPrimersProduct length
Alpha-A fusionAlpha-linker + linker-AAlphaL+SacI + AP+NotI 1500 bp
Omega-A fusionOmega-linker + linker-AOmegaL+SacI + AP+NotI 1440 bp

PCL program for fusion A with alpha and omega

TemperatureTime
94°C4:00
94°C0:3028 cycles
48°C-62°C0:45
72°C2:30
72°C10:00
4°Cinfinite

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