Team:Warsaw/Calendar-Main/3 October 2008

From 2008.igem.org

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<li> Planting on LB with kanamycin.</li></ol>
<li> Planting on LB with kanamycin.</li></ol>
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<h3>Preparation of pACYC177 + OmpA_omega_</h3>
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<h4>Piotr</h4>
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<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 pACYC177 + OmpA_omega_.</li>
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<li> Planting on LB with kanamycin.</li></ol>
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<h4>Piotr</h4>
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<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 ligations <a href=http://partsregistry.org/Part:pSB2K3>pSB2K3</a> + _alpha, <a href=http://partsregistry.org/Part:pSB2K3>pSB2K3</a> + _omega and pACYC177 + OmpA_omega_ (both digested with BamHI and PstI - from 30 September).</li>
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<li> Planting on LB with kanamycin.</li></ol>
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Revision as of 23:25, 26 October 2008

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Preparation of pSB2K3 + _alpha

Piotr

  1. Transformation of TOP10 with ligation pSB2K3 + linker_alpha (BBa_K103009).
  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 pACYC177 + OmpA_omega_

Piotr

  1. Transformation of TOP10 with ligation pACYC177 + OmpA_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).

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