Team:Warsaw/Calendar-Main/27 August 2008

From 2008.igem.org

(Difference between revisions)
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 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 (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> <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>
Line 26: Line 26:
 +
<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>

Revision as of 10:08, 26 October 2008

Gallery Bricks Notebook Team Project Home


Previous day
return to main notebook page
Previous entry
next notebook entry

 


Tests for ampicillin resistance given by protein added to medium

Piotr

Results of growth from previus 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. <img src=" Go29.jpg" width=300/> Fig. 1. Control NdeI/NotI digest of isolated plasmid pGeneart+A
    1. Marker
    2. digested plasmid pGeneart+A