Team:Warsaw/Calendar-Main/26 June 2008

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<h3><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#pcl">Polymerase Chain Ligation (PCL)</a></h3><h4>Michał L., Ewa, Marcin</h4>
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<h3>Preparation of alpha-A and omega-A fusions</h3><h4>Michał L., Ewa, Marcin</h4>
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<b><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#pcl">Polymerase Chain Ligation (PCL)</a></b>
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Revision as of 12:12, 26 October 2008

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Change of the reporter from pZC320 with B-galactosidase to GFP or RFP

Piotr, Weronika

  1. Isolation of pZC30
  2. Inoculation of E.coli TOP10 carrying pSB1A2 standard plasmids carrying parts: BBa_E0840(Gfp genetrator) and BBa_J04450 (RFP generator)

Preparation of constructs with OmpA protein fusions

Michał K.

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

Preparation of alpha-A and omega-A fusions

Michał L., Ewa, Marcin

Polymerase Chain Ligation (PCL)
ProductTemplatesPrimersProduct length
Alpha-A fusionAlpha-linker + linker-AAlphaL+SacI + AP+NotI 1100 bp
Omega-A fusionOmega-linker + linker-AOmegaL+SacI + AP+NotI 750 bp


PCL program for fusion A with alpha and omega
TemperatureTime
94°C4:00
94°C0:3028 cycles
48°C-62°C0:45
68°C1:30
68°C10:00
4°Cinfinite

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