Team:Warsaw/Calendar-Main/17 July 2008

From 2008.igem.org

(Difference between revisions)
Line 9: Line 9:
<li> Gel electrophoresis 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 (420 bp - pKS-A lane, 4050 bp - pACYC177+OmpA_omega lane and 4300 bp pACYC177+OmpA_alpha lane). </li>
<li> Gel electrophoresis 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 (420 bp - pKS-A lane, 4050 bp - pACYC177+OmpA_omega lane and 4300 bp pACYC177+OmpA_alpha lane). </li>
<li> <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#ligation">Ligation</a> of A DNA fragment with both pACYC177 vectors.</li>
<li> <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#ligation">Ligation</a> of A DNA fragment with both pACYC177 vectors.</li>
-
<li> Transformation of <i>E. coli</i> <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top">TOP10</a>  strain with ligations. </li>
+
<li> Transformation of <i>E. coli</i> <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">TOP10</a>  strain with ligations. </li>
<li>Transformants plating on LB + kanamycin. </li></ol>
<li>Transformants plating on LB + kanamycin. </li></ol>
</p>
</p>
Line 16: Line 16:
<p><ol><li><a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#digest">Digest</a> of pET15b-OmpA-omega and Z(in GeneArt vector) with NdeI and NotI.</li>  
<p><ol><li><a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#digest">Digest</a> of pET15b-OmpA-omega and Z(in GeneArt vector) with NdeI and NotI.</li>  
<li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#DNA_isolation_from_agarose_gel">Gel-out</a> of Z (~200 bp band).</li>
<li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#DNA_isolation_from_agarose_gel">Gel-out</a> of Z (~200 bp band).</li>
-
<li>Overnight<a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#ligation">Ligation</a> of Z into digested pET15b-OmpA-omega.</li>
+
<li>Overnight <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#ligation">Ligation</a> of Z into digested pET15b-OmpA-omega.</li>

Revision as of 16:51, 5 October 2008

Gallery Bricks Notebook Team Project Home


Previous day
return to main notebook page
Previous entry
Interactive list of topics
next notebook entry

 



Cloning of protein A DNA to OmpA constructs

  1. Digest of pKS-A plasmid, pACYC177+OmpA_omega and pACYC177+OmpA_alpha with NotI and SacI (pACYC177 were also dephosporylated with CIAP)
  2. Gel electrophoresis and gel-out of proper bands (420 bp - pKS-A lane, 4050 bp - pACYC177+OmpA_omega lane and 4300 bp pACYC177+OmpA_alpha lane).
  3. Ligation of A DNA fragment with both pACYC177 vectors.
  4. Transformation of E. coli TOP10 strain with ligations.
  5. Transformants plating on LB + kanamycin.

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

  1. Digest of pET15b-OmpA-omega and Z(in GeneArt vector) with NdeI and NotI.
  2. Gel-out of Z (~200 bp band).
  3. Overnight Ligation of Z into digested pET15b-OmpA-omega.

Retrieved from "http://2008.igem.org/Team:Warsaw/Calendar-Main/17_July_2008"