Team:Warsaw/Calendar-Main/17 October 2008

From 2008.igem.org

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<h3>Preparation of pT7_alpha_link</h3>
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<h3>Preparation of <a href=http://partsregistry.org/wiki/index.php?title=Part:pSB1A3>pSB1A3</a> + <a href=http://partsregistry.org/Part:BBa_K103019>alpha_linker under PT7 (BBa_K103019)</a></h3>
<h4>Michał K., Piotr</h4>
<h4>Michał K., Piotr</h4>
<ol>
<ol>
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<li>Gel electrophoresis of over night digest (<A href=http://partsregistry.org/wiki/index.php?title=Part:pSB1A3>pSB1A3</a> carrying  pT7_alpha) and <a href="https://2008.igem.org/Wiki/Team:Warsaw/protocols#DNA_isolation_from_agarose_gel">gel-out</a> of proper band - 800 bp. </li>
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<li>Gel electrophoresis of over night digest (<A href=http://partsregistry.org/wiki/index.php?title=Part:pSB1A3>pSB1A3</a> carrying  <a href=http://partsregistry.org/Part:BBa_K103019>alpha_linker under PT7 (BBa_K103019)</a>) and <a href="https://2008.igem.org/Wiki/Team:Warsaw/protocols#DNA_isolation_from_agarose_gel">gel-out</a> of proper band - 800 bp. </li>
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<li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#ligation">Ligation</a> of isolated DNA fragments: <a href=http://partsregistry.org/wiki/index.php?title=Part:pSB2K3>pSB2K3</a> (from 30 September) + pT7_alpha.</li>
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<li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#ligation">Ligation</a> of isolated DNA fragments: <a href=http://partsregistry.org/wiki/index.php?title=Part:pSB2K3>pSB2K3</a> (from 30 September) + <a href=http://partsregistry.org/Part:BBa_K103019>alpha_linker under PT7 (BBa_K103019)</a>.</li>
<li> <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#chemotransform">Transformation</a> of <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">TOP10</a> with above ligation.</li>
<li> <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#chemotransform">Transformation</a> of <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">TOP10</a> with above ligation.</li>
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<li> Planting on LB with kanamycin.</li></ol>
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<li> Plating on LB with kanamycin.</li></ol>

Revision as of 10:12, 27 October 2008

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Preparation of pACYC177 + OmpA-linker-omega-linker (BBa_K103016)

Michał K.

  1. Isolation of plasmids from cultures inoculated on previous day pACYC177 + OmpA-linker-omega-linker (BBa_K103016).
  2. Control digest of isolated plasmids with EcoRI and PstI (Orange buffer). Gel electrophoresis - no proper clones found.

Preparation of pSB1A3 + alpha_linker under PT7 (BBa_K103019)

Michał K., Piotr

  1. Gel electrophoresis of over night digest (pSB1A3 carrying alpha_linker under PT7 (BBa_K103019)) and gel-out of proper band - 800 bp.
  2. Ligation of isolated DNA fragments: pSB2K3 (from 30 September) + alpha_linker under PT7 (BBa_K103019).
  3. Transformation of TOP10 with above ligation.
  4. Plating on LB with kanamycin.

Piotr

  1. Transformation of TOP10 with overnight ligation - RFP(?????) + AraC+pBAD+AID and 5kb(?????) + pT7_alpha.
  2. Planting: RFP(?????) + AraC+pBAD+AID on LB with ampicillin and 5kb(?????) + pT7_alpha on LB with kanamycin.

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