Team:Warsaw/Calendar-Main/22 July 2008

From 2008.igem.org

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<ol>
<ol>
<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>
<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 (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 plasmids with SacI and NotI (BamHI buffer).</li>
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<li>Gel electrophoresis of productsof digest.</li>
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<li>Gel electrophoresis of products of digest.</li>
<li>2 selected clones were send to DNA sequencing lab</li>
<li>2 selected clones were send to DNA sequencing lab</li>
</ol>
</ol>
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<h3>Cloning of protein A DNA to OmpA constructs</h3><h4>Michał K.</h4>
<h3>Cloning of protein A DNA to OmpA constructs</h3><h4>Michał K.</h4>
<p><ol>
<p><ol>
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<li>Colony PCR on colonies from plates with transformations pACYC177+OmpA_A_omega and pACYC177+OmpA_A_alpha. Primers used:
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<li>Colony <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#pcr">PCR</a> on colonies from plates with transformations pACYC177+OmpA_A_omega and pACYC177+OmpA_A_alpha. Primers used:
<a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#AL+SacI">AL+SacI</a> and <a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#AP+NotI">AP+NotI.</a>
<a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#AL+SacI">AL+SacI</a> and <a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#AP+NotI">AP+NotI.</a>

Revision as of 18:58, 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 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 products of digest.
  4. 2 selected clones were send to DNA sequencing lab

Cloning of protein A DNA to OmpA constructs

Michał K.

  1. Colony PCR on colonies from plates with transformations pACYC177+OmpA_A_omega and pACYC177+OmpA_A_alpha. Primers used: AL+SacI and AP+NotI.
  2. Confirmed transformant colonies (found only on pACYC177+OmpA_A_alpha plate) inoculated to liquid LB with kanamycin.

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