Team:Warsaw/Calendar-Main/7 August 2008

From 2008.igem.org

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<p><ol>
<p><ol>
<li>Isolation of plasmids from cultures inoculated on previous day.</li>
<li>Isolation of plasmids from cultures inoculated on previous day.</li>
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<li>Control <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest">digest</a> of isolated plasmids (<a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-omega-deltaA>pACYC177+omega_&Delta;A</a>) with HindIII and SacI (BamHI buffer)(Fig. 1.). 7 from 8 colonies confirmed. </li></ol></p>
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<li>Control <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest">digest</a> of isolated plasmids (<a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-omega-deltaA>pACYC177+omega_&Delta;A</a>) with HindIII and SacI (BamHI buffer)<a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/7_August_2008#fig1">Fig. 1</a>. 7 from 8 colonies confirmed. </li></ol></p>
<h3>Checking whether degradation of the fusions with OmpA is caused by Lon and OmpT proteases (present in <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">TOP10</a>)</h3>
<h3>Checking whether degradation of the fusions with OmpA is caused by Lon and OmpT proteases (present in <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">TOP10</a>)</h3>
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<img src="https://static.igem.org/mediawiki/2008/8/89/Zz.jpg" width=300/> <var><b>Fig. 1. Control SacI/HindIII digests of isolated plasmids</b><br>  
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<a name="fig1"><img src="https://static.igem.org/mediawiki/2008/8/89/Zz.jpg" width=300/></a> <var><b>Fig. 1. Control SacI/HindIII digests of isolated plasmids</b><br>  
1. Marker<br>
1. Marker<br>
2-9. digested plasmids pACYC177+OmpA_omega_deltaA<br></var>
2-9. digested plasmids pACYC177+OmpA_omega_deltaA<br></var>

Revision as of 17:47, 26 October 2008

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Purification of proteins: Z-alpha and Z-omega

Piotr, Emilia

pET15b+Z-alpha and pET15b+Z-omega inoculated for overnight culture (Rosetta strain)

Preparing of pACYC177+OmpA_omega_ΔA construct

Michał K.

  1. Isolation of plasmids from cultures inoculated on previous day.
  2. Control digest of isolated plasmids (pACYC177+omega_ΔA) with HindIII and SacI (BamHI buffer)Fig. 1. 7 from 8 colonies confirmed.

Checking whether degradation of the fusions with OmpA is caused by Lon and OmpT proteases (present in TOP10)

Michał L.

Separate transformant colonies (Rosetta transformation from previous day) inoculated to liquid LB with kanamycin.

Fig. 1. Control SacI/HindIII digests of isolated plasmids
1. Marker
2-9. digested plasmids pACYC177+OmpA_omega_deltaA

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