Team:Warsaw/Calendar-Main/25 July 2008

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<h3>Cloning of protein Z DNA to pET15b-OmpA-omega in place of OmpA</h3>
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<h3>Cloning of protein Z DNA to <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pET15b%2BOmpA-omega>pET15b-OmpA-omega</a> in place of OmpA</h3>
<h4>Paweł</h4>
<h4>Paweł</h4>
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<p><ol>
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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 plasmids from cultures inocluated on previous day</li>
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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 plasmids from cultures inoculated on previous day.</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 NdeI and NotI. Two positives obtained (proper band: ~200 bp visible)</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 NdeI and NotI. Two positives obtained (proper band: ~200 bp visible).</li>
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<li>One of positive plasmids transformed into <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">TOP10</a> and plated on LB+amp, overnight, for further isolation of pET15b+Z+omega vector</li>
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<li>One of positive plasmids transformed into <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">TOP10</a> and plated on LB+amp, overnight, for further isolation of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pET15b%2Bhis%2BZ%2Bomega>pET15b+Z+omega</a> vector.</li>
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<h3>Cloning of protein A DNA to OmpA constructs</h3>
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<h3>Cloning of protein A DNA to OmpA constructs</h3><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 of plasmids</a> from cultures inocluated on previous day (pACYC177+OmpA_A_omega).</li>
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<ol><li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#plasmid_DNA_isolation">Isolation of plasmids</a> from cultures inocluated on previous day (<a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-A-omega>pACYC177+OmpA_A_omega</a>).</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 BamHI and SacI (BamHI buffer). Proper clone founded. </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 with BamHI and SacI (BamHI buffer).</li><li> Gel electrophoresis - proper clone found. </li></ol>
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<ol><li>Separate transformant colonies (tranformation of ligation of pACYC177+OmpA_alpha and omega_A from previous day) inoculated to liquid LB with kanamycin.
 
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</li></ol>
 
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Cloning of protein Z DNA to pET15b-OmpA-omega in place of OmpA

Paweł

  1. Isolation of plasmids from cultures inoculated on previous day.
  2. Control digest of isolated plasmids with NdeI and NotI. Two positives obtained (proper band: ~200 bp visible).
  3. One of positive plasmids transformed into TOP10 and plated on LB+amp, overnight, for further isolation of pET15b+Z+omega vector.

Cloning of protein A DNA to OmpA constructs

Michał K.

  1. Isolation of plasmids from cultures inocluated on previous day (pACYC177+OmpA_A_omega).
  2. Control digest of isolated plasmids with BamHI and SacI (BamHI buffer).
  3. Gel electrophoresis - proper clone found.


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