Team:Warsaw/Calendar-Main/29 August 2008

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<p><ol><li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#plasmid_DNA_isolation">Isolation</a> of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pET15b%2BA-alpha>pET15b+A-alpha</a> plasmids.</li>
<p><ol><li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#plasmid_DNA_isolation">Isolation</a> of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pET15b%2BA-alpha>pET15b+A-alpha</a> plasmids.</li>
<li> Control <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#digest">digest</a> of isolated plasmids with XbaI and BamHI (Tango buffer). </li>
<li> Control <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#digest">digest</a> of isolated plasmids with XbaI and BamHI (Tango buffer). </li>
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<li>Gel electrophoresis (Fig. 1.). Choice of proper clone. </li>
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<li>Gel electrophoresis <a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/29_August_2008#fig1">Fig. 1</a>. Choice of proper clone. </li>
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<img src="
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<a name="fig1"><img src="
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https://static.igem.org/mediawiki/2008/a/a8/Kokodak.jpg" width=300/> <var><b>Fig. 1. Control XbaI/BamHI digests of  plasmids isolated from transformants with colony PCR product.</b><br>  
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https://static.igem.org/mediawiki/2008/a/a8/Kokodak.jpg" width=300/></a> <var><b>Fig. 1. Control XbaI/BamHI digests of  plasmids isolated from transformants with colony PCR product.</b><br>  
1. Marker<br>
1. Marker<br>
2-4. digested plasmids pET15b+A-alpha <br></var>
2-4. digested plasmids pET15b+A-alpha <br></var>

Revision as of 17:59, 26 October 2008

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

Michał K.

  1. Isolation of pET15b+A-alpha plasmids.
  2. Control digest of isolated plasmids with XbaI and BamHI (Tango buffer).
  3. Gel electrophoresis Fig. 1. Choice of proper clone.

Purification of proteins: A-alpha

Piotr

Single transformations of Rosetta with plasmids pET15b+A-alpha. Plating on LB with chloramphenicol and ampicillin.

Blue-white screening and rifampicin test in GM2163

Michał L.

GM2163 competent cells carrying pZC320 were transformed with following plasmids:

Transformants were selected on CTAXI plates (LB + Chloramphenicol + Tetracycline + IPTG + Ampicillin 30 μg/mL + X-Gal) because GM2163 are chloramphenicol resistant.

Fig. 1. Control XbaI/BamHI digests of plasmids isolated from transformants with colony PCR product.
1. Marker
2-4. digested plasmids pET15b+A-alpha



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