Team:Warsaw/Calendar-Main/1 July 2008

From 2008.igem.org

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<h3>Change of the reporter from pZC with B-galactosidaze to GFP or RFP</h3>
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<h3>Change of the reporter from <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pZC320>pZC320</a> with B-galactosidase to GFP or RFP</h3>
<h4>Piotr, Weronika</h4>
<h4>Piotr, Weronika</h4>
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<li>Gel electrophoresis of digested DNA - there where no proper plasmids</li>
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<li>Isolation of pZC</li>
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<li>Isolation of <A href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pZC320>pZC320</a></li>
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<li>Isolation of nazwa (Gfp genetrator), nazwa (RFP generator)</li>
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<li>Isolation of <a href=http://partsregistry.org/wiki/index.php?title=Part:pSB1A2>pSB1A2</a> standard plasmids carrying parts: <A href=http://partsregistry.org/wiki/index.php?title=Part:BBa_E0840>BBa_E0840</a>(Gfp genetrator) and <a href=http://partsregistry.org/wiki/index.php?title=Part:BBa_J04450>BBa_J04450</a> (RFP generator)</li>
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<li>Owernight digest of nazwa (Gfp genetrator), nazwa (RFP generator) with NotI</li>
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<li>Overnight digest of <a href=http://partsregistry.org/wiki/index.php?title=Part:pSB1A2>pSB1A2</a> standard plasmids carrying parts: <A href=http://partsregistry.org/wiki/index.php?title=Part:BBa_E0840>BBa_E0840</a>(Gfp genetrator) and <a href=http://partsregistry.org/wiki/index.php?title=Part:BBa_J04450>BBa_J04450</a> (RFP generator) with NotI</li>
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Owernight digest of pZC with NotI and defosforylation with CIAP
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Overnight digest of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pZC320>pZC320</a> with NotI and <a href=https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#removing>dephosphorylation</a> with CIAP
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Revision as of 16:35, 24 October 2008

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

Piotr, Weronika

  1. Isolation of plasmids from transformants.
  2. Control digest of plasmids with NotI.
  3. Gel electrophoresis of digested DNA - there where no proper plasmids
  4. Isolation of pZC320
  5. Isolation of pSB1A2 standard plasmids carrying parts: BBa_E0840(Gfp genetrator) and BBa_J04450 (RFP generator)
  6. Overnight digest of pSB1A2 standard plasmids carrying parts: BBa_E0840(Gfp genetrator) and BBa_J04450 (RFP generator) with NotI
  7. Overnight digest of pZC320 with NotI and dephosphorylation with CIAP

Cloning of alpha-A and omega-A fusions on pKS

Michał L., Ewa, Marcin

  1. Isolation of plasmids from transformants.
  2. Control digest of plasmids with SacI+NotI (BamHI buffer).
  3. Gel electrophoresis of digested DNA.

We have successfully cloned A-alpha and A-omega fusions on pKS vector. The DNA will be now sequenced to ensure that our constructs are correct.

Preparation of constructs with OmpA protein fusions

Paweł

  1. Isolation of pET15b plasmid.
  2. Overnight digest of pET15b plasmid with NdeI and BamHI (Tango 2x buffer), dephosphorylation with CIAP.