Team:Warsaw/Calendar-Main/16 July 2008

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<h3>Cloning omega-A fusion on <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pKSII%2B>pKS</a> (second attempt)</h3>
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<h4>Michał L., Ewa, Marcin</h4>
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<p>We have massive outbreak of some virulent bacteriophage strain in our lab. All our current E. coli cultures are gone :-(. Until we get rid of the phage there will be no microbiological work in our lab. Sorry.<p/>
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<h3>Cloning of protein Z DNA to OmpA constructs</h3><h4>Michał K.</h4>
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<h3>Cloning of protein Z DNA to OmpA constructs<br>Michał K.</h3>
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<li> Isolation of plasmids from cultures inocluated on previous day (pACYC177+OmpA_Z_alpha). </li>
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<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-Z-omega>pACYC177+OmpA_Z_omega</a>). </li>
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<li> Control digest of isolated plasmids with BamHI and SacI (we found 4 good clones).</li></ol></p>
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<li> Control  
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<a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#digest">digest</a> of isolated plasmids with BamHI and SacI (BamHI buffer).</li> <li> Gel electrophoresis - both checked colones are good (<a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/16_July_2008#fig1">Fig. 1.</a>).</li>
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<a name="fig1"><img src="https://static.igem.org/mediawiki/2008/a/ac/Dwa.jpg" width=300/></a> <var><b>Fig. 1. </b>Control SacI/BamHI digests of isolated plasmids<br>
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<h3>Cloning of protein A DNA to geneart plasmid in place of protein Z DNA<br>Antoni</h3>
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1. Marker<br>
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2-3. digested plasmids pACYC177+OmpA_Z_omega <br></var>
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<li> <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#electrotransform">Transformation</a> of <i>E. coli</i> <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top">TOP10</a> strain with ligation from previous day </li>
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Latest revision as of 17:55, 26 October 2008

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Cloning omega-A fusion on pKS (second attempt)

Michał L., Ewa, Marcin

We have massive outbreak of some virulent bacteriophage strain in our lab. All our current E. coli cultures are gone :-(. Until we get rid of the phage there will be no microbiological work in our lab. Sorry.

Cloning of protein Z DNA to OmpA constructs

Michał K.

  1. Isolation of plasmids from cultures inocluated on previous day (pACYC177+OmpA_Z_omega).
  2. Control digest of isolated plasmids with BamHI and SacI (BamHI buffer).
  3. Gel electrophoresis - both checked colones are good (Fig. 1.).

Fig. 1. Control SacI/BamHI digests of isolated plasmids
1. Marker
2-3. digested plasmids pACYC177+OmpA_Z_omega

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