Team:Warsaw/Calendar-Main/16 October 2008

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<li>Control <a href=https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest>digest</a> of isolated plasmids: 3kb(?????)+ link_alpha, 3kb(????)_pLac_OmpA_ + alpha2, RFP(?????) + AraC+pBAD+AID and RFP(?????) + pT7_alpha (14 October) with EcoRI and PstI (Orange buffer). Gel electrophoresis - no proper clones found for RFP(?????) + AraC+pBAD+AID ligation but we found good clones for other ligations.</li>
<li>Control <a href=https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest>digest</a> of isolated plasmids: 3kb(?????)+ link_alpha, 3kb(????)_pLac_OmpA_ + alpha2, RFP(?????) + AraC+pBAD+AID and RFP(?????) + pT7_alpha (14 October) with EcoRI and PstI (Orange buffer). Gel electrophoresis - no proper clones found for RFP(?????) + AraC+pBAD+AID ligation but we found good clones for other ligations.</li>
<li> Inoculation of colonies from plate with ligation of pACYC177 + OmpA_omega_ to LB with kanamycin. </li>
<li> Inoculation of colonies from plate with ligation of pACYC177 + OmpA_omega_ to LB with kanamycin. </li>
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<li> Repetition of <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#ligation">ligation</a> of isolated DNA fragments: RFP(?????) + AraC+pBAD+AID.</li>
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<li> Repetition of overnight <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#ligation">ligation</a> of isolated DNA fragments: RFP(?????) + AraC+pBAD+AID.</li>
<li>Overnight <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#digest">digest</a> of RFP(?????) + pT7_alpha EcoRI and BcuI (BamHI buffer). </li>
<li>Overnight <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#digest">digest</a> of RFP(?????) + pT7_alpha EcoRI and BcuI (BamHI buffer). </li>
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<h3></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 above ligation.</li>
 
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<li> Planting on LB with ampicillin.</li></ol>
 
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Revision as of 19:53, 25 October 2008

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Preparation of BioBricks

Michał K.

  1. Isolation of plasmids from culture inoculated on previous day 3kb(?????)+ link_alpha, 3kb(????)_pLac_OmpA_ + alpha2 and RFP(?????) + AraC+pBAD+AID.
  2. Control digest of isolated plasmids: 3kb(?????)+ link_alpha, 3kb(????)_pLac_OmpA_ + alpha2, RFP(?????) + AraC+pBAD+AID and RFP(?????) + pT7_alpha (14 October) with EcoRI and PstI (Orange buffer). Gel electrophoresis - no proper clones found for RFP(?????) + AraC+pBAD+AID ligation but we found good clones for other ligations.
  3. Inoculation of colonies from plate with ligation of pACYC177 + OmpA_omega_ to LB with kanamycin.
  4. Repetition of overnight ligation of isolated DNA fragments: RFP(?????) + AraC+pBAD+AID.
  5. Overnight digest of RFP(?????) + pT7_alpha EcoRI and BcuI (BamHI buffer).

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