Team:Warsaw/Calendar-Main/27 August 2008

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<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>
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2. digested plasmid pGeneart+A<br></var>
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Revision as of 10:09, 26 October 2008

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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. Fig. 1. Control NdeI/NotI digest of isolated plasmid pGeneart+A
    1. Marker
    2. digested plasmid pGeneart+A




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