Team:Warsaw/Calendar-Main/27 August 2008

From 2008.igem.org

(Difference between revisions)
 
(4 intermediate revisions not shown)
Line 4: Line 4:
<html>
<html>
<h3>Tests for ampicillin resistance given by protein added to medium</h3><h4>Piotr</h4>
<h3>Tests for ampicillin resistance given by protein added to medium</h3><h4>Piotr</h4>
-
Results of growth from previus day
+
Results of growth from previous day
<br><br>
<br><br>
<table id="result">
<table id="result">
Line 19: Line 19:
<ol><li> <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#digest">Digest</a> of <A href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pGeneart-A>pGeneart+A</a> and <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pET15b%2BOmpA-alpha>pET15b+OmpA-alpha</a> with NdeI and NotI (Orange buffer), pET15b was also <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#removing"> dephosphorylated</a>. </li>  
<ol><li> <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#digest">Digest</a> of <A href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pGeneart-A>pGeneart+A</a> and <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pET15b%2BOmpA-alpha>pET15b+OmpA-alpha</a> with NdeI and NotI (Orange buffer), pET15b was also <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#removing"> dephosphorylated</a>. </li>  
-
<li> Gel electrophoresis of digested plasmids (Fig. 1.) 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 A lane and 6200 bp for pET15b lane).  </li>
+
<li> Gel electrophoresis of digested plasmids (<a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/27_August_2008#fig1">Fig. 1.</a>) 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 A 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 <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pET15b%2BA-alpha>pET15b-Alpha and A</a>. </li>
<li> <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#ligation">Ligation</a> of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pET15b%2BA-alpha>pET15b-Alpha and A</a>. </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> 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>
<li> Transformants plating on LB + ampicillin.</li>
 +
 +
<a name="fig1"><img src="
 +
https://static.igem.org/mediawiki/2008/1/1b/Go29.jpg" width=300/></a> <var><b>Fig. 1.</b> NdeI/NotI digest of plasmid pGeneart+A<br>
 +
1. Marker<br>
 +
2. Digested plasmid pGeneart+A<br></var>
 +
</html>
</html>
-
<img src="
+
 
-
https://static.igem.org/mediawiki/2008/1/1b/Go29.jpg" width=300/> <var><b>Fig. 1. Control NdeI/NotI digest of isolated plasmid pGeneart+A</b><br>
+
-
1. Marker<br>
+
-
2. digested plasmid pGeneart+A<br></var>
+

Latest revision as of 20:56, 26 October 2008

Gallery Bricks Notebook Team Project Home


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

 


Tests for ampicillin resistance given by protein added to medium

Piotr

Results of growth from previous day

OmpA variant ("hunter")"Prey" variantGrowth with preyGrowth without prey
OmpA-alphaHis+Z+omega+-
OmpA-alphaHis+Z+alfa--
OmpA-A-alphaHis+Z+omega+-

Cloning of protein A DNA to pET15b+OmpA-alpha plasmid

Michał K.

  1. Digest of pGeneart+A and pET15b+OmpA-alpha with NdeI and NotI (Orange buffer), pET15b was also dephosphorylated.
  2. Gel electrophoresis of digested plasmids (Fig. 1.) and gel-out of proper bands (400 bp for A lane and 6200 bp for pET15b lane).
  3. Ligation of pET15b-Alpha and A.
  4. Transformation of E. coli TOP10.
  5. Transformants plating on LB + ampicillin.
    6. Fig. 1. NdeI/NotI digest of plasmid pGeneart+A
    1. Marker
    2. Digested plasmid pGeneart+A




Retrieved from "http://2008.igem.org/Team:Warsaw/Calendar-Main/27_August_2008"