Team:Warsaw/Calendar-Main/25 July 2008

From 2008.igem.org

(Difference between revisions)
 
(13 intermediate revisions not shown)
Line 4: Line 4:
<html>
<html>
-
<p>1. Isolation of plasmids from cultures inocluated on previous day.<br>
 
-
2. Control digest of isolated plasmids with BamHI and SacI (we confirmed 4 colonies but A in our constructs turned out to be trunctated and contain only one from two highly similar domains). 
 
-
</p>
 
-
 
+
<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>
-
<br>
+
<h4>Paweł</h4>
-
<h3>Cloning of protein A DNA to pET15b+OmpA-alfa plasmid in place of OmpA<br>Antoni</h3>
+
<p><ol>
<p><ol>
-
<li> Colony PCR on colonies from plates with transformations pGeneart+A Primers used:
+
<li><a href=https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#plasmid_DNA_isolation>Isolation</a> of plasmids from cultures inoculated on previous day.</li>
 +
<li>Control <a href=https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest>digest</a> of isolated plasmids with NdeI and NotI. Two positives obtained (proper band: ~200 bp visible).</li>
 +
<li>One of positive plasmids transformed into <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">TOP10</a> and plated on LB+amp, overnight, for further isolation of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pET15b%2Bhis%2BZ%2Bomega>pET15b+Z+omega</a> vector.</li>
 +
</ol></p>
 +
 
 +
 
 +
 
 +
<h3>Cloning of protein A DNA to OmpA constructs</h3><h4>Michał K.</h4>
 +
<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 previous day (<a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-A-omega>pACYC177+OmpA_A_omega</a>).</li>
 +
<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 (BamHI buffer).</li><li> Gel electrophoresis - proper clone found. </li></ol>
 +
 
 +
 
-
<a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#AP+NotI">AP+NotI</a> and <a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#AL+SacI">AL+SacI</a>
 
-
<li> Lack confirmed transformant colonies </li>
 
</html>
</html>
 +
<!-- do not remove this! -->
<!-- do not remove this! -->
{{WarNotebookEnd}}
{{WarNotebookEnd}}

Latest revision as of 14:52, 25 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 Z DNA to pET15b-OmpA-omega in place of OmpA

Paweł

  1. Isolation of plasmids from cultures inoculated on previous day.
  2. Control digest of isolated plasmids with NdeI and NotI. Two positives obtained (proper band: ~200 bp visible).
  3. One of positive plasmids transformed into TOP10 and plated on LB+amp, overnight, for further isolation of pET15b+Z+omega vector.

Cloning of protein A DNA to OmpA constructs

Michał K.

  1. Isolation of plasmids from cultures inocluated on previous day (pACYC177+OmpA_A_omega).
  2. Control digest of isolated plasmids with BamHI and SacI (BamHI buffer).
  3. Gel electrophoresis - proper clone found.


Retrieved from "http://2008.igem.org/Team:Warsaw/Calendar-Main/25_July_2008"