Team:Warsaw/Calendar-Main/29 August 2008

From 2008.igem.org

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<h3>Cloning of protein A DNA to pET15b+Omp-alpha plasmid</h3><h4>Michał K.</h4>
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<h3>Cloning of protein A DNA to <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pET15b%2BOmpA-alpha>pET15b+Omp-alpha</a> plasmid</h3><h4>Michał K.</h4>
<p><ol><li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#plasmid_DNA_isolation">Isolation</a> of pET15b+A-alpha plasmids.</li>
<p><ol><li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#plasmid_DNA_isolation">Isolation</a> of pET15b+A-alpha plasmids.</li>
<li> Control <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#digest">digest</a> of isolated plasmids with NdeI and NotI (Orange buffer). </li>
<li> Control <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#digest">digest</a> of isolated plasmids with NdeI and NotI (Orange buffer). </li>
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<h4>Piotr</h4>
<h4>Piotr</h4>
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Single transformations of <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#rosetta">Rosetta</a> with plasmids pET15b+A-alfa, pET15b+Z-alfa and pET15b+Z-omega. Plating them on LB with chloramphenicol and ampicillin.</p></html>
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Single transformations of <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#rosetta">Rosetta</a> with plasmids pET15b+A-alpha, pET15b+Z-alpha and pET15b+Z-omega. Plating them on LB with chloramphenicol and ampicillin.</p></html>

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Cloning of protein A DNA to pET15b+Omp-alpha plasmid

Michał K.

  1. Isolation of pET15b+A-alpha plasmids.
  2. Control digest of isolated plasmids with NdeI and NotI (Orange buffer).
  3. Gel elctrophoresis. Chioce of proper clone.

Protein purification: A-alpha, Z-alpha i Z-omega

Piotr

Single transformations of Rosetta with plasmids pET15b+A-alpha, pET15b+Z-alpha and pET15b+Z-omega. Plating them on LB with chloramphenicol and ampicillin.