Team:Warsaw/Calendar-Main/8 October 2008

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<h3>Preparation of <a href=http://partsregistry.org/Part:pSB2K3>pSB2K3</a> + _alpha</h3>
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<h3>Preparation of <a href=http://partsregistry.org/Part:pSB2K3>pSB2K3</a> + <a href=http://partsregistry.org/Part:BBa_K103009>linker_alpha (BBa_K103009)</a></h3>
<h4>Michał K., Piotr</h4>
<h4>Michał K., Piotr</h4>
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<li><a href=https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#plasmid_DNA_isolation>Isolation</a> of plasmid from culture inoculated on previous day (<a href=http://partsregistry.org/Part:pSB2K3>pSB2K3</a> + _alpha).</li>
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<li><a href=https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#plasmid_DNA_isolation>Isolation</a> of plasmid from culture inoculated on previous day (<a href=http://partsregistry.org/Part:pSB2K3>pSB2K3</a> + <a href=http://partsregistry.org/Part:BBa_K103009>linker_alpha (BBa_K103009)</a>).</li>
<li>Control <a href=https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest>digest</a> of isolated plasmid with EcoRI and PstI (Orange buffer). Gel electrophoresis - still no proper clones found.</li>
<li>Control <a href=https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest>digest</a> of isolated plasmid with EcoRI and PstI (Orange buffer). Gel electrophoresis - still no proper clones found.</li>
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<li> Tranformants planting on LB with tetracycline. </li>
<li> Tranformants planting on LB with tetracycline. </li>
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<li><a href=https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#plasmid_DNA_isolation>Isolation</a> of plasmid from culture inoculated on previous day (<a href=http://partsregistry.org/Part:pSB2K3>pSB2K3</a> + _alpha, <a href=http://partsregistry.org/Part:pSB2K3>pSB2K3</a> + _omega and pACYC177 + OmpA_omega_).</li>
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<li><a href=https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#plasmid_DNA_isolation>Isolation</a> of plasmid from culture inoculated on previous day (<a href=http://partsregistry.org/Part:pSB2K3>pSB2K3</a> + <a href=http://partsregistry.org/Part:BBa_K103009>linker_alpha (BBa_K103009)</a>, <a href=http://partsregistry.org/Part:pSB2K3>pSB2K3</a> + _omega and pACYC177 + OmpA_omega_).</li>
<li>Control <a href=https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest>digest</a> of isolated plasmid with EcoRI and PstI (Orange buffer). Gel electrophoresis - proper clones found for <a href=http://partsregistry.org/Part:pSB2K3>pSB2K3</a> + _omega.</li>
<li>Control <a href=https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest>digest</a> of isolated plasmid with EcoRI and PstI (Orange buffer). Gel electrophoresis - proper clones found for <a href=http://partsregistry.org/Part:pSB2K3>pSB2K3</a> + _omega.</li>

Revision as of 23:26, 26 October 2008

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Preparation of pSB2K3 + 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 - still no proper clones found.

Preparation of pSB2K3 + _omega

Piotr

  1. Isolation of plasmid from culture inoculated on previous day (pSB2K3 + _omega).
  2. Control digest of isolated plasmid with EcoRI and PstI (Orange buffer). Gel electrophoresis - proper clones found.

Preparation of BioBricks

Michał K.

  1. Colony PCR with AIDlNrH and AIDpLinB primers on colonies from plates with transformations pSB1A3+ AID (annealing temperature - 55°C, 60 s of elongation step).
  2. Inoculation of confirmed colonies to liquid LB + ampicillin.
  3. Inoculation of colonies from plate with ligation of pET15b+OmpA_omega without XbaI to liquid LB + ampicillin.

Piotr

  1. Inoculation of colonies from plate with ligation of pSB2K3 + pLac_OmpA_omega (without EcoRI site) to liquid LB + kanamycin.
  2. Transformation of TOP10 with ligation pMPMT5+AID without EcoRI site.
  3. Tranformants planting on LB with tetracycline.
  4. Isolation of plasmid from culture inoculated on previous day (pSB2K3 + linker_alpha (BBa_K103009), pSB2K3 + _omega and pACYC177 + OmpA_omega_).
  5. Control digest of isolated plasmid with EcoRI and PstI (Orange buffer). Gel electrophoresis - proper clones found for pSB2K3 + _omega.

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