Team:Warsaw/Calendar-Main/3 October 2008

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<h3>Preparation of <a href=http://partsregistry.org/Part:pSB2K3>pSB2K3</a> + <a href=http://partsregistry.org/wiki/index.php?title=Part:BBa_K103009>linker_alpha (BBa_K103009)</h3>
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<h3>Preparation of <a href=http://partsregistry.org/Part:pSB2K3>pSB2K3</a> + _alpha</h3>
<h4>Piotr</h4>
<h4>Piotr</h4>
<ol><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 ligation <a href=http://partsregistry.org/Part:pSB2K3>pSB2K3</a> + _alpha.</li>
<ol><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 ligation <a href=http://partsregistry.org/Part:pSB2K3>pSB2K3</a> + _alpha.</li>

Revision as of 22:41, 26 October 2008

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Preparation of pSB2K3 + _alpha

Piotr

  1. Transformation of TOP10 with ligation pSB2K3 + _alpha.
  2. Planting on LB with kanamycin.

Preparation of pSB2K3 + _omega

Piotr

  1. Transformation of TOP10 with ligation pSB2K3 + _omega.
  2. Planting on LB with kanamycin.

Preparation of BioBricks

Michał K.

  1. Isolation of plasmid from culture inoculated on previous day (pSB1A3+Omp_).
  2. Control digest of isolated plasmid with EcoRI and PstI (Orange buffer). Gel electrophoresis - proper clones found.
  3. Digest of pET15b+OmpA_omega with XbaI (Tango buffer). DNA ends blunting with Klenow fragment (3 hr).
  4. Gel electrophoresis and gel-out of proper band (pET15b+OmpA_omega without XbaI - 6500 bp).

Piotr

  1. Transformation of TOP10 with ligations pSB2K3 + _alpha, pSB2K3 + _omega and pACYC177 + OmpA_omega_ (both digested with BamHI and PstI - from 30 September).
  2. Planting on LB with kanamycin.

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