Team:Warsaw/Calendar-Main/26 August 2008

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Revision as of 22:00, 12 October 2008


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Colony PCR on colonies from plates with transformations pGeneart+A Primers used: AP+NotI and AL+SacI.
Lack of confirmed transformant colonies.

Digest pET15b+OmpA_alpha and pGeneart+Z with NdeI and NotI (Orange buffer), pET15b+OmpA_alpha dephosphorylation with CIAP.
Gel electrophoresis of digested plasmids and gel-out of proper bands (200 bp for Z lane and 6200 bp for pET15b lane).
Ligation of pET15b+alpha and Z.
Transformation of E. coli TOP10.
Transformants plating on LB + ampicillin.

@@ Line 11: / Line 11: @@
 <li> Lack of confirmed transformant colonies. </li></ol></p>
-<h3>Cloning of protein Z DNA to pET15b+Omp-alfa plasmid</h3><h4>Antoni</h4>
+<h3>Cloning of protein Z DNA to pET15b+Omp-alpha plasmid</h3><h4>Antoni</h4>
 <p><ol>
 <li> <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#digest">Digest</a> pET15b+OmpA_alpha and pGeneart+Z with NdeI and NotI (Orange buffer), pET15b+OmpA_alpha<a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#removing"> dephosphorylation with CIAP</a>. </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 (200 bp for Z lane and 6200 bp for pET15b lane).  </li>
-<li> <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#ligation">Ligation</a> of pET15b+alfa and Z. </li>
+<li> <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#ligation">Ligation</a> of pET15b+alpha and Z. </li>
 <li> Transformation of <i>E. coli</i> <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">TOP10</a>. </li>
 <li> Transformants plating on LB + ampicillin.</li>