Team:Warsaw/Calendar-Main/7 October 2008

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<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 ligations (3kb??????) + pLac_OmpA_omega (without EcoRI site) and pET15b+OmpA_omega without XbaI.</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 ligations (3kb??????) + pLac_OmpA_omega (without EcoRI site) and pET15b+OmpA_omega without XbaI.</li>
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<li> Planting: (3kb??????) + pLac_OmpA_omega (without EcoRI site) on LB with kanamycin and pET15b+OmpA_omega without XbaI on LB with ampicillin.</li></ol>
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<li> Planting: (3kb??????) + pLac_OmpA_omega (without EcoRI site) on LB with kanamycin and pET15b+OmpA_omega without XbaI on LB with ampicillin.</li>
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<li>Inoculation of few more colonies which grown on plates with three separate transformations: (3kb??????) + _alpha, (3kb????) + _omega and pACYC177 + OmpA_omega_ to liquid LB + kanamycin. </li>
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Revision as of 11:18, 25 October 2008

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Preparation of BioBricks

Michał K.

  1. Isolation of plasmid from culture inoculated on previous day ((3kb??????) + _alpha, (3kb????) + _omega and pACYC177 + OmpA_omega_).
  2. Control digest of isolated plasmid with EcoRI and PstI (Orange buffer). Gel electrophoresis - no proper clones found.
  3. Overnight ligation of DNA fragments: RFP(????)+ AID.

Piotr

  1. Transformation of TOP10 with ligations (3kb??????) + pLac_OmpA_omega (without EcoRI site) and pET15b+OmpA_omega without XbaI.
  2. Planting: (3kb??????) + pLac_OmpA_omega (without EcoRI site) on LB with kanamycin and pET15b+OmpA_omega without XbaI on LB with ampicillin.
  3. Inoculation of few more colonies which grown on plates with three separate transformations: (3kb??????) + _alpha, (3kb????) + _omega and pACYC177 + OmpA_omega_ to liquid LB + kanamycin.


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