Team:Warsaw/Calendar-Main/28 August 2008

From 2008.igem.org

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<h3>Cloning of protein Z DNA to pET15b+Omp-alpha plasmid</h3><h4>Antoni</h4>
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<h3>Cloning of protein Z DNA to <A href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pET15b%2BOmpA-alpha>pET15b+Omp-alpha</a> plasmid</h3><h4>Antoni</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+Z-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+Z-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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<li>Gel elctrophoresis. Chioce of proper clone. </li>
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<li>Gel electrophoresis. Choice of proper clone. </li>
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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>
<ol><li> Colony <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#pcr">PCR</a> on colonies from plates with transformation pEt15b+A_alpha. <br>Primers used:
<ol><li> Colony <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#pcr">PCR</a> on colonies from plates with transformation pEt15b+A_alpha. <br>Primers used:

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

Antoni

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

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

Michał K.

  1. Colony PCR on colonies from plates with transformation pEt15b+A_alpha.
    Primers used: AP+NotI and AL+SacI.
  2. Gel electrophoresis.
  3. Confirmed transformant colonies inoculated to liquid LB with ampicillin.

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