Team:Warsaw/Calendar-Main/16 October 2008

From 2008.igem.org

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<h3>Preparation of pT7_alpha_link</h3>
<h3>Preparation of pT7_alpha_link</h3>
<h4>Michał K.</h4>
<h4>Michał K.</h4>
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<p>Control <a href=https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest>digest</a> of isolated plasmid - <A href=http://partsregistry.org/wiki/index.php?title=Part:pSB1A3>pSB1A3</a> carrying  pT7_alpha fragment (14 October) with EcoRI and PstI (Orange buffer). Gel electrophoresis - proper clones found.</p>
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<ol><li>Control <a href=https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest>digest</a> of isolated plasmid - <A href=http://partsregistry.org/wiki/index.php?title=Part:pSB1A3>pSB1A3</a> carrying  pT7_alpha fragment (14 October) with EcoRI and PstI (Orange buffer). Gel electrophoresis - proper clones found.</li>
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<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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<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> 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>
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<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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Revision as of 09:13, 27 October 2008

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Preparation of pSB2K3 + linker_alpha (BBa_K103009)

Michał K.

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

Preparation of pACYC177 + OmpA-linker-omega-linker (BBa_K103016)

Michał K.

Inoculation of colonies from plate with new ligation of pACYC177 + OmpA-linker-omega-linker (BBa_K103016) to LB with kanamycin.

Preparation of pT7_alpha_link

Michał K.

  1. Control digest of isolated plasmid - pSB1A3 carrying pT7_alpha fragment (14 October) with EcoRI and PstI (Orange buffer). Gel electrophoresis - proper clones found.
  2. Overnight digest of RFP(?????) + pT7_alpha EcoRI and BcuI (BamHI buffer).

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. Repetition of overnight ligation of isolated DNA fragments: RFP(?????) + AraC+pBAD+AID.

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