Team:Warsaw/Calendar-Main/18 July 2008

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

Paweł

Ligation transformed into TOP10 strain and plated on LB + ampicillin.

Cloning of protein Z DNA to OmpA constructs

Michał K.

Colonies from transformation pACYC177+OmpA_Z_alpha were inoculated to liquid LB broth with kanamycin.

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-<h3>Cloning of protein A DNA to OmpA constructs</h3>
-<p><ol>
-<li>Colony PCR 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>
+<h3>Cloning of protein Z DNA to <A href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pET15b%2BOmpA-omega>pET15b-OmpA-omega</a> in place of OmpA</h3><h4>Paweł</h4>
-</li>
-<li> Confirmed transformant colonies (found only on pACYC177+OmpA_A_alpha plate) inoculated to liquid LB with kanamycin. </li></ol>
-</p>
-<h3>Paweł</h3>
-<p><ol><li>Ligation transformed into TOP10 strain and plated on LB+kanamycin</li>
-</ol></p>
-<br>
+<p>Ligation <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#chemotransform">transformed</a> into <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">TOP10</a> strain and plated on LB + ampicillin.</p>
-<h3>Cloning of protein A DNA to pET15b+Omp-alfa plasmid<br>Antoni</h3>
-<p><ol>
-<li> Isolation of plasmids from cultures inocluated on previous day (pGeneart+A and pET15b+OmpA-alfa). </li>
-<li> <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#digest">Digest</a> pET15b+OmpA_alpha and pGeneart+A with NdeI and NotI, pET15b+OmpA_alpha<a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#removing"> dephosphorylation with CIAP</a> </li> </li>
-<li> Gel electrophoresis of digested plasmids and <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#DNA_isolation_from_agarose_gel">gel-out</a> of proper bands  </li>
-<li> Overnight <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#ligation">ligation</a> of pET15b+alfa and A </li>
-</li>
-</ol></p>
+<h3>Cloning of protein Z DNA to OmpA constructs</h3>
+<h4>Michał K.</h4>
+<p>Colonies from transformation <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-Z-alpha>pACYC177+OmpA_Z_alpha</a> were inoculated to liquid LB broth with kanamycin. </p>
+<br>