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Team:Warsaw/Calendar-Main/29 August 2008
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<h3>Cloning of protein A DNA to <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pET15b%2BOmpA-alpha>pET15b+Omp-alpha</a> plasmid</h3><h4>Michał K.</h4> | <h3>Cloning of protein A DNA to <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pET15b%2BOmpA-alpha>pET15b+Omp-alpha</a> plasmid</h3><h4>Michał K.</h4> | ||
<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 | + | <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>Gel elctrophoresis. Chioce of proper clone. </li> | <li>Gel elctrophoresis. Chioce of proper clone. </li> | ||
</ol></p> | </ol></p> | ||
Revision as of 14:03, 25 October 2008
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Cloning of protein A DNA to pET15b+Omp-alpha plasmidMichał K.
Protein purification: A-alphaPiotrSingle transformations of Rosetta with plasmids pET15b+A-alpha. Plating on LB with chloramphenicol and ampicillin. Blue-white screening and rifampicin test in GM2163Michał 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.
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