Team:Warsaw/Calendar-Main/7 October 2008

From 2008.igem.org

(Difference between revisions)
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</ol>
</ol>
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<h3>Preparation of pACYC177 + OmpA_omega_</h3>
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<h4>Michał K., Piotr</h4>
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<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 plasmid from culture inoculated on previous day (pACYC177 + OmpA_omega_).</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 plasmid with EcoRI and PstI (Orange buffer). Gel electrophoresis - no proper clones found.</li>
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<li>Inoculation of few more colonies which grown on plate with transformation: pACYC177 + OmpA_omega_ to liquid LB + kanamycin. </li></ol>
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<h4>Michał K.</h4>
<h4>Michał K.</h4>
<ol>
<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 plasmid from culture inoculated on previous day (<a href=http://partsregistry.org/Part:pSB2K3>pSB2K3</a> + _alpha, <a href=http://partsregistry.org/Part:pSB2K3>pSB2K3</a> + _omega and pACYC177 + OmpA_omega_).</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 plasmid with EcoRI and PstI (Orange buffer). Gel electrophoresis - no proper clones found.</li>
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<li> Overnight <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#ligation">ligation</a> of DNA fragments: <a href=http://partsregistry.org/wiki/index.php?title=Part:pSB1A3>pSB1A3</a>+ AID.</li>
<li> Overnight <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#ligation">ligation</a> of DNA fragments: <a href=http://partsregistry.org/wiki/index.php?title=Part:pSB1A3>pSB1A3</a>+ AID.</li>
</ol>
</ol>
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<ol><li> <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#chemotransform">Transformation</a> of <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">TOP10</a> with ligations <a href=http://partsregistry.org/Part:pSB2K3>pSB2K3</a> + pLac_OmpA_omega (without EcoRI site) and <a hrefhttps://2008.igem.org/Wiki/Team:Warsaw/vectors/pET15b%2BOmpA-omega>pET15b+OmpA_omega</a> without XbaI.</li>
<ol><li> <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#chemotransform">Transformation</a> of <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">TOP10</a> with ligations <a href=http://partsregistry.org/Part:pSB2K3>pSB2K3</a> + pLac_OmpA_omega (without EcoRI site) and <a hrefhttps://2008.igem.org/Wiki/Team:Warsaw/vectors/pET15b%2BOmpA-omega>pET15b+OmpA_omega</a> without XbaI.</li>
<li> Plating: <a href=http://partsregistry.org/Part:pSB2K3>pSB2K3</a> + pLac_OmpA_omega (without EcoRI site) on LB with kanamycin and pET15b+OmpA_omega without XbaI on LB with ampicillin.</li>
<li> Plating: <a href=http://partsregistry.org/Part:pSB2K3>pSB2K3</a> + pLac_OmpA_omega (without EcoRI site) on LB with kanamycin and pET15b+OmpA_omega without XbaI on LB with ampicillin.</li>
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<li>Inoculation of few more colonies which grown on plates with three separate transformations: <a href=http://partsregistry.org/Part:pSB2K3>pSB2K3</a> + _alpha, <a href=http://partsregistry.org/Part:pSB2K3>pSB2K3</a> + _omega and pACYC177 + OmpA_omega_ to liquid LB + kanamycin. </li>
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<li><a href="https://2008.igem.org/Wiki/Team:Warsaw/protocols#digest">Digest</a> of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5-AID>pMPMT5+AID</a> with EcoRI (EcoRI buffer). DNA ends <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#blunting">blunting</a> with Klenow fragment. </li>
<li><a href="https://2008.igem.org/Wiki/Team:Warsaw/protocols#digest">Digest</a> of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5-AID>pMPMT5+AID</a> with EcoRI (EcoRI buffer). DNA ends <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#blunting">blunting</a> with Klenow fragment. </li>
<li> Overnight <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#ligation">ligation</a> of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5-AID>pMPMT5+AID</a> without EcoRI site.</li>
<li> Overnight <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#ligation">ligation</a> of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5-AID>pMPMT5+AID</a> without EcoRI site.</li>

Revision as of 23:33, 26 October 2008

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Preparation of pSB2K3 + linker_alpha (BBa_K103009)

Michał K., Piotr

  1. Isolation of plasmid from culture inoculated on previous day (pSB2K3 + linker_alpha (BBa_K103009)).
  2. Control digest of isolated plasmid with EcoRI and PstI (Orange buffer). Gel electrophoresis - no proper clones found.
  3. Inoculation of few more colonies which grown on plate with transformation: pSB2K3 + linker_alpha (BBa_K103009) to liquid LB + kanamycin.

Preparation of pSB2K3 + _omega

Michał K., Piotr

  1. Isolation of plasmid from culture inoculated on previous day (pSB2K3 + _omega).
  2. Control digest of isolated plasmid with EcoRI and PstI (Orange buffer). Gel electrophoresis - no proper clones found.
  3. Inoculation of few more colonies which grown on plates with transformation: pSB2K3 + _omega to liquid LB + kanamycin.

Preparation of pACYC177 + OmpA_omega_

Michał K., Piotr

  1. Isolation of plasmid from culture inoculated on previous day (pACYC177 + OmpA_omega_).
  2. Control digest of isolated plasmid with EcoRI and PstI (Orange buffer). Gel electrophoresis - no proper clones found.
  3. Inoculation of few more colonies which grown on plate with transformation: pACYC177 + OmpA_omega_ to liquid LB + kanamycin.

Preparation of BioBricks

Michał K.

  1. Overnight ligation of DNA fragments: pSB1A3+ AID.

Piotr

  1. Transformation of TOP10 with ligations pSB2K3 + pLac_OmpA_omega (without EcoRI site) and pET15b+OmpA_omega without XbaI.
  2. Plating: pSB2K3 + pLac_OmpA_omega (without EcoRI site) on LB with kanamycin and pET15b+OmpA_omega without XbaI on LB with ampicillin.
  3. Digest of pMPMT5+AID with EcoRI (EcoRI buffer). DNA ends blunting with Klenow fragment.
  4. Overnight ligation of pMPMT5+AID without EcoRI site.

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