Team:Warsaw/Calendar-Main/7 October 2008

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(Difference between revisions)
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<h3>Preparation of <a href=http://partsregistry.org/Part:BBa_K103018>OmpA_linker_omega_linker under Plac (BBa_K103018)</a></h3>
<h3>Preparation of <a href=http://partsregistry.org/Part:BBa_K103018>OmpA_linker_omega_linker under Plac (BBa_K103018)</a></h3>
<h4>Piotr</h4>
<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> + <a href=http://partsregistry.org/Part:BBa_K103018>BBa_K103018</a> (without EcoRI site).</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 <a href=http://partsregistry.org/Part:pSB2K3>pSB2K3</a> + <a href=http://partsregistry.org/Part:BBa_K103018>BBa_K103018</a> (without internal EcoRI site).</li>
<li> Plating on LB with kanamycin.</li>
<li> Plating on LB with kanamycin.</li>

Revision as of 20:14, 27 October 2008

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Preparation of linker_alpha (BBa_K103009)

Michał K., Piotr

  1. Isolation of plasmid from culture inoculated on previous day (pSB2K3 + linker_alpha (BBa_K103009)).
  2. Control digest of isolated plasmid with EcoRI and PstI (Orange buffer). Gel electrophoresis - no proper clones found.
  3. Inoculation of few more colonies which grown on plate with transformation: pSB2K3 + linker_alpha (BBa_K103009) to liquid LB + kanamycin.

Preparation of linker_omega (BBa_K103013)

Michał K., Piotr

  1. Isolation of plasmid from culture inoculated on previous day (pSB2K3 + linker_omega (BBa_K103013)).
  2. Control digest of isolated plasmid with EcoRI and PstI (Orange buffer). Gel electrophoresis - no proper clones found.
  3. Inoculation of few more colonies which grown on plates with transformation: pSB2K3 + linker_omega (BBa_K103013) to liquid LB + kanamycin.

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

Michał K., Piotr

  1. Isolation of plasmid from culture inoculated on previous day (pACYC177 + OmpA-linker-omega-linker (BBa_K103016)).
  2. Control digest of isolated plasmid with EcoRI and PstI (Orange buffer). Gel electrophoresis - no proper clones found.
  3. Inoculation of few more colonies which grown on plate with transformation: pACYC177 + OmpA-linker-omega-linker (BBa_K103016) to liquid LB + kanamycin.

Preparation of vector for pT7 constructs

Piotr

  1. Transformation of TOP10 with selfligation of pET15b+OmpA_omega without XbaI.
  2. Plating on LB with ampicillin.

Preparation of OmpA_linker_omega_linker under Plac (BBa_K103018)

Piotr

  1. Transformation of TOP10 with ligations pSB2K3 + BBa_K103018 (without internal EcoRI site).
  2. Plating on LB with kanamycin.

Preparation of AID

Michał K., Piotr

  1. Ligation of DNA fragments: pSB1A3+ AID (from 1 October).
  2. Transformation of TOP10 with above ligation.
  3. Plating on LB with ampicillin.

Preparation of AraC+pBAD+AID

Piotr

  1. Digest of pMPMT5+AID with EcoRI (EcoRI buffer). DNA ends blunting with Klenow fragment.
  2. Overnight ligation of pMPMT5+AID without EcoRI site.

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