Team:Warsaw/Calendar-Main/22 August 2008

From 2008.igem.org

(Difference between revisions)
Line 6: Line 6:
<html>
<html>
-
<h3>Cloning of protein A DNA to pET15b+Omp-alfa plasmid</h3><h4>Antoni</h4>
+
<h3>Cloning of protein A DNA to pET15b+Omp-alpha plasmid</h3><h4>Antoni</h4>
<p><ol>
<p><ol>
-
<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 <a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/21_August_2008">previous day</a> (pGeneart+A and pET15b+OmpA-alfa). </li>
+
<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 <a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/21_August_2008">previous day</a> (pGeneart+A and pET15b+OmpA-alpha). </li>
<li> Control <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#digest">digest</a> of pGeneart+A plasmids with NdeI and NotI (Orange buffer). </li>
<li> Control <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#digest">digest</a> of pGeneart+A plasmids with NdeI and NotI (Orange buffer). </li>
<li>Gel elctrophoresis. Chioce of proper clone. </li>
<li>Gel elctrophoresis. Chioce of proper clone. </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 (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>
<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 (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>
<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 (400 bp for protein A lane and 6200 for pET15b lane). </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 (400 bp for protein A lane and 6200 for pET15b lane). </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> Overnight <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#ligation">ligation</a> of pET15b+alpha and A. </li>
</ol></p>
</ol></p>
</html>
</html>

Revision as of 21:58, 12 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 pET15b+Omp-alpha plasmid

Antoni

  1. Isolation of plasmids from cultures inocluated on previous day (pGeneart+A and pET15b+OmpA-alpha).
  2. Control digest of pGeneart+A plasmids with NdeI and NotI (Orange buffer).
  3. Gel elctrophoresis. Chioce of proper clone.
  4. Digest pET15b+OmpA_alpha and pGeneart+A with NdeI and NotI (Orange buffer), pET15b+OmpA_alpha dephosphorylation with CIAP.
  5. Gel electrophoresis of digested plasmids and gel-out of proper bands (400 bp for protein A lane and 6200 for pET15b lane).
  6. Overnight ligation of pET15b+alpha and A.


Retrieved from "http://2008.igem.org/Team:Warsaw/Calendar-Main/22_August_2008"