Team:Warsaw/Calendar-Main/22 July 2008

From 2008.igem.org

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<h3>Cloning of protein Z DNA to pET15b-OmpA-omega in place of OmpA<br>
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<h3>Cloning of protein Z DNA to pET15b-OmpA-omega in place of OmpA</h3>
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Paweł</h3>
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<h4>Paweł</h4>
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<p><ol><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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<p><ol><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>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 (Orange buffer). Zero positive results obtained - just empty vectors</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 (Orange buffer). Zero positive results obtained - just empty vectors.</li>
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<li>Another overnight <a href=https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#ligation>ligation</a> of pET15b-OmpA-omega (NdeI/NotI cutted) with protein Z DNA</li>
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<li>Another overnight <a href=https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#ligation>ligation</a> of pET15b-OmpA-omega (NdeI/NotI cutted) with protein Z DNA.</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>
<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>
<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>
<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. </li></ol>
<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. </li></ol>
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<h4>Michał L., Ewa, Marcin:</h4>
<h4>Michał L., Ewa, Marcin:</h4>
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<li>Isolation of plasmids from transformants</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 transformants</li>
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<li>Digest of plasmids with SacI and NotI</li>
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<li>Digest of plasmids with SacI and NotI (BamHI buffer).</li>
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<li>Gel electrophoresis of products</li>
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<li>Gel electrophoresis of productsof digest.</li>
<li>2 selected clones were send to DNA sequencing lab</li>
<li>2 selected clones were send to DNA sequencing lab</li>
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Revision as of 18:51, 11 October 2008

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Cloning of protein Z DNA to pET15b-OmpA-omega in place of OmpA

Paweł

  1. Isolation of plasmids from cultures inocluated on previous day.
  2. Control digest of isolated plasmids with NdeI and NotI (Orange buffer). Zero positive results obtained - just empty vectors.
  3. Another overnight ligation of pET15b-OmpA-omega (NdeI/NotI cutted) with protein Z DNA.

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.

Cloning omega-A fusion on pKS (second attempt)

Michał L., Ewa, Marcin:

  1. Isolation of plasmids from transformants
  2. Digest of plasmids with SacI and NotI (BamHI buffer).
  3. Gel electrophoresis of productsof digest.
  4. 2 selected clones were send to DNA sequencing lab


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