Team:Warsaw/Calendar-Main/7 August 2008

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<h3>Purification of proteins: Z-alpha and Z-omega</h3><h4>Piotr, Emilia</h4>
<h3>Purification of proteins: Z-alpha and Z-omega</h3><h4>Piotr, Emilia</h4>
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<p><a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pET15b%2BZ-alpha>pET15b+Z-alpha</a> and <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pET15b%2Bhis%2BZ%2Bomega>pET15b+Z-omega</a> inoculated for overnight culture (Rosetta strain)</p>
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<p><a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pET15b%2BZ-alpha>pET15b+Z-alpha</a> and <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pET15b%2Bhis%2BZ%2Bomega>pET15b+Z-omega</a> inoculated for overnight culture (Rosetta strain).</p>
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<h3>Preparing of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-omega-deltaA>pACYC177+OmpA_omega_&Delta;A</a> construct</h3><h4>Michał K.</h4>
 
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<p><ol>
 
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<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)<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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<p>Separate transformant colonies (Rosetta transformation from previous day) inoculated to liquid LB with kanamycin.</p>
<p>Separate transformant colonies (Rosetta transformation from previous day) inoculated to liquid LB with kanamycin.</p>
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<h3>Preparing <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-omega-deltaA>pACYC177+OmpA_omega_&Delta;A</a> construct</h3><h4>Michał K.</h4>
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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>  
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<p><ol>
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<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)(<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>
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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. </b>Control SacI/HindIII digests of isolated plasmids<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>

Latest revision as of 18:35, 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).

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.

Preparing 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.

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

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