Team:Warsaw/Calendar-Main/23 July 2008
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- | <h3>Cloning | + | <h3>Cloning omega-A fusion on <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pKSII%2B>pKS</a> (second attempt)</h3> |
- | + | <h4>Michał L., Ewa, Marcin</h4> | |
- | < | + | <ol> |
- | <li><a href=https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#plasmid_DNA_isolation>Isolation</a> of plasmids from | + | <li><a href=https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#plasmid_DNA_isolation>Isolation</a> of plasmids from transformants.</li> |
- | <li> | + | <li><a href=https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest>Digest</a> of plasmids with SacI and NotI (BamHI buffer).</li> |
- | <li> | + | <li>Gel electrophoresis of products of digest.</li> |
- | < | + | <li>2 selected clones were send to DNA sequencing lab.</li> |
- | + | </ol> | |
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+ | <h3>Cloning of protein Z DNA to <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pET15b%2BOmpA-omega>pET15b-OmpA-omega</a> in place of OmpA</h3><h4>Paweł</h4> | ||
+ | <p>Ligations <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#chemotransform">transformed</a> into <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">TOP10</a> and plated on LB + ampicillin.</p> | ||
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+ | <h3>Preparation of chemocompetent bacteria</h3> | ||
+ | <h4>Antoni</h4> | ||
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- | + | Setup of overnight culture <i>E. coli</i> <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">TOP10</a> (LB broth) for preparation of chemocompetent bacteria.</p> | |
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+ | <h3>Cloning of protein A DNA to OmpA constructs</h3><h4>Michał K.</h4> | ||
+ | <ol> | ||
+ | <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-A-alpha>pACYC177+OmpA_A_alpha</a>). </li> | ||
+ | <li>Control <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest">digest</a> of isolated plasmids with BamHI and SacI (BamHI buffer).</li><li> Gel electrophoresis - proper clone found (<a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/23_July_2008#fig1">Fig. 1.</a>).</li></ol> | ||
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+ | <a name="fig1"><img src="https://static.igem.org/mediawiki/2008/f/fb/23_july.jpg" width=300/></a> <var><b>Fig. 1. </b>Control SacI/BamHI digests of pACYC177+OmpA_A_alpha<br> | ||
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+ | 1-4. digested plasmids isolated from transformants obtained previous day<br> | ||
+ | 5. Marker<br></var> | ||
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Latest revision as of 21:23, 29 October 2008
Cloning omega-A fusion on pKS (second attempt)Michał L., Ewa, Marcin
Cloning of protein Z DNA to pET15b-OmpA-omega in place of OmpAPawełLigations transformed into TOP10 and plated on LB + ampicillin. Preparation of chemocompetent bacteriaAntoniSetup of overnight culture E. coli TOP10 (LB broth) for preparation of chemocompetent bacteria. Cloning of protein A DNA to OmpA constructsMichał K.
1-4. digested plasmids isolated from transformants obtained previous day 5. Marker
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