Team:Warsaw/Calendar-Main/16 October 2008

From 2008.igem.org

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<h3>Preparation of AraC+pBAD+AID</h3>
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<h4>Michał K.</h4>
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<ol><li><a href=https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#plasmid_DNA_isolation>Isolation</a> of plasmids from culture inoculated on previous day (<a href=http://partsregistry.org/wiki/index.php?title=Part:pSB1A3>pSB1A3</a> + AraC+pBAD+AID).</li>
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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 with EcoRI and PstI (Orange buffer). Gel electrophoresis - still no proper clones found.</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: <a href=http://partsregistry.org/wiki/index.php?title=Part:pSB1A3>pSB1A3</a> + AraC+pBAD+AID.</li></ol>
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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 (<a href=https://2008.igem.org/Team:Warsaw/Calendar-Main/14_October_2008>14 October</a>) 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 (<a href=https://2008.igem.org/Team:Warsaw/Calendar-Main/14_October_2008>14 October</a>) 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>
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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>
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Revision as of 11:31, 27 October 2008

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Preparation of 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 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 alpha_linker under PT7 (BBa_K103019)

Michał K.

  1. Control digest of isolated plasmid - pSB1A3 carrying alpha_linker under PT7 (BBa_K103019) fragment (14 October) with EcoRI and PstI (Orange buffer). Gel electrophoresis - proper clones found.
  2. Overnight digest pSB1A3 carrying alpha_linker under PT7 (BBa_K103019) with EcoRI and BcuI (BamHI buffer).

Preparation of AraC+pBAD+AID

Michał K.

  1. Isolation of plasmids from culture inoculated on previous day (pSB1A3 + AraC+pBAD+AID).
  2. Control digest of isolated plasmids with EcoRI and PstI (Orange buffer). Gel electrophoresis - still no proper clones found.
  3. Repetition of overnight ligation of isolated DNA fragments: pSB1A3 + AraC+pBAD+AID.

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.

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