Team:Warsaw/Calendar-Main/8 October 2008

From 2008.igem.org

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<h4>Michał K.</h4>
<h4>Michał K.</h4>
<p>Inoculation of colonies from plate with ligation of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pET15b%2BOmpA-omega>pET15b+OmpA_omega</a> without XbaI to liquid LB + ampicillin. </p>
<p>Inoculation of colonies from plate with ligation of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pET15b%2BOmpA-omega>pET15b+OmpA_omega</a> without XbaI to liquid LB + ampicillin. </p>
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<h3>Preparation of pLac_OmpA_omega</h3>
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<h4>Piotr</h4>
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<p> Inoculation of colonies from plate with ligation of <a href=http://partsregistry.org/Part:pSB2K3>pSB2K3</a> + pLac_OmpA_omega (without EcoRI site) to liquid LB + kanamycin.</p>
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<h3></h3>
<h3></h3>
<h4>Piotr</h4>
<h4>Piotr</h4>
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<ol><li> Inoculation of colonies from plate with ligation of <a href=http://partsregistry.org/Part:pSB2K3>pSB2K3</a> + pLac_OmpA_omega (without EcoRI site) to liquid LB + kanamycin.</li>
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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 <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5-AID>pMPMT5+AID</a> without EcoRI site.</li>
<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=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5-AID>pMPMT5+AID</a> without EcoRI site.</li>

Revision as of 10:43, 27 October 2008

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Preparation of linker_alpha (BBa_K103009)

Piotr

  1. Isolation of plasmid from culture inoculated on previous day (pSB2K3 + linker_alpha (BBa_K103009)).
  2. Control digest of isolated plasmids with EcoRI and PstI (Orange buffer). Gel electrophoresis - still no proper clones found.

Preparation of linker_omega (BBa_K103013)

Piotr

  1. Isolation of plasmid from culture inoculated on previous day (pSB2K3 + linker_omega (BBa_K103013)).
  2. Control digest of isolated plasmids with EcoRI and PstI (Orange buffer). Gel electrophoresis - proper clones found.

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

Piotr

  1. Isolation of plasmid from culture inoculated on previous day (pACYC177 + OmpA-linker-omega-linker (BBa_K103016)).
  2. Control digest of isolated plasmids with EcoRI and PstI (Orange buffer). Gel electrophoresis - no proper clones found.

Preparation of vector for pT7 constructs

Michał K.

Inoculation of colonies from plate with ligation of pET15b+OmpA_omega without XbaI to liquid LB + ampicillin.

Preparation of pLac_OmpA_omega

Piotr

Inoculation of colonies from plate with ligation of pSB2K3 + pLac_OmpA_omega (without EcoRI site) to liquid LB + kanamycin.

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.

Piotr

  1. Transformation of TOP10 with ligation pMPMT5+AID without EcoRI site.
  2. Tranformants planting on LB with tetracycline.

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