Team:Warsaw/Calendar-Main/9 October 2008

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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 (RFP(?????)+ AID and pACYC177 + pET15b+OmpA_omega without XbaI).</li>
<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 (RFP(?????)+ AID and pACYC177 + pET15b+OmpA_omega without XbaI).</li>
<li>Control <a href=https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest>digest</a> of isolated RFP(?????)+ AID plasmids with EcoRI and PstI (Orange buffer) and pET15b+OmpA_omega without XbaI plasmids with XbaI and BamHI (Tango buffer). Gel electrophoresis - proper clones found for both ligations.</li>
<li>Control <a href=https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest>digest</a> of isolated RFP(?????)+ AID plasmids with EcoRI and PstI (Orange buffer) and pET15b+OmpA_omega without XbaI plasmids with XbaI and BamHI (Tango buffer). Gel electrophoresis - proper clones found for both ligations.</li>
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<li><a href=https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest>Digest</a> of pET15b+OmpA_omega without XbaI plasmid with NdeI and SacI (BamHI buffer). </li>
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<li><a href=https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest>Digest</a> of pET15b+OmpA_omega without XbaI plasmid with NdeI and SacI (BamHI buffer), <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#removing">dephosphorylation</a> with CIAP </li>
<li>Gel electrophoresis and <a href="https://2008.igem.org/Wiki/Team:Warsaw/protocols#DNA_isolation_from_agarose_gel">gel-out</a> of proper band: ??????. </li>
<li>Gel electrophoresis and <a href="https://2008.igem.org/Wiki/Team:Warsaw/protocols#DNA_isolation_from_agarose_gel">gel-out</a> of proper band: ??????. </li>

Revision as of 12:59, 25 October 2008

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Preparation of BioBricks

Michał K.

  1. Isolation of plasmid from culture inoculated on previous day (RFP(?????)+ AID and pACYC177 + pET15b+OmpA_omega without XbaI).
  2. Control digest of isolated RFP(?????)+ AID plasmids with EcoRI and PstI (Orange buffer) and pET15b+OmpA_omega without XbaI plasmids with XbaI and BamHI (Tango buffer). Gel electrophoresis - proper clones found for both ligations.
  3. Digest of pET15b+OmpA_omega without XbaI plasmid with NdeI and SacI (BamHI buffer), dephosphorylation with CIAP
  4. Gel electrophoresis and gel-out of proper band: ??????.

Piotr

  1. Isolation of plasmid from culture inoculated on previous day (3kb??????) + pLac_OmpA_omega (without EcoRI site).
  2. Control digest of isolated (3kb??????) + pLac_OmpA_omega with EcoRI and PstI (Orange buffer) proper clones found.
  3. Inoculation of colonies from plate with ligation of pMPMT5+AID without EcoRI site to liquid LB + tetracycline.

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