Team:Warsaw/Calendar-Main/21 July 2008

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<p>'''Cloning of protein A DNA to OmpA constructs''' <br>
 
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1. Colony PCR on colonies from plates with transformations pACYC177+OmpA_A_omega. Primers used:
 
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<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>
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<h3>Cloning of protein A DNA to OmpA constructs</h3>
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<ol><li>Colony PCR on colonies from plates with transformations pACYC177+OmpA_A_omega. Primers used:
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2. Confirmed transformant colonies inoculated to liquid LB with kanamycin. <br>
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3. <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_alpha). <br>  
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<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></li>
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4. 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 .
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<li>Confirmed transformant colonies inoculated to liquid LB with kanamycin. </li>
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<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_alpha). </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 BamHI and SacI.</li></ol>
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<h3>Cloning of protein Z DNA to pET15b-OmpA-omega in place of OmpA<br>
<h3>Cloning of protein Z DNA to pET15b-OmpA-omega in place of OmpA<br>
Paweł</h3>
Paweł</h3>
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<p><ol><li>Ligations transformed into <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">TOP10</a> and plated on LB + ampicillin</li></ol></p>
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<ol><li>Ligations transformed into <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">TOP10</a> and plated on LB + ampicillin.</li></ol>
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Revision as of 17:05, 5 October 2008

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Cloning of protein A DNA to OmpA constructs

  1. Colony PCR on colonies from plates with transformations pACYC177+OmpA_A_omega. Primers used: AL+SacI and AP+NotI
  2. Confirmed transformant colonies inoculated to liquid LB with kanamycin.
  3. Isolation of plasmids from cultures inocluated on previous day (pACYC177+OmpA_A_alpha).
  4. Control digest of isolated plasmids with BamHI and SacI.

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

  1. Ligations transformed into TOP10 and plated on LB + ampicillin.

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