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Team:Warsaw/Calendar-Main/29 August 2008
From 2008.igem.org
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<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 XbaI and BamHI (Tango buffer). </li> | <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 electrophoresis. Choice of proper clone. </li> | + | <li>Gel electrophoresis (Fig. 1.). Choice of proper clone. </li> |
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<p>Transformants were selected on CTAXI plates (LB + Chloramphenicol + Tetracycline + IPTG + Ampicillin 30 μg/mL + X-Gal) because GM2163 are chloramphenicol resistant.</p> | <p>Transformants were selected on CTAXI plates (LB + Chloramphenicol + Tetracycline + IPTG + Ampicillin 30 μg/mL + X-Gal) because GM2163 are chloramphenicol resistant.</p> | ||
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| + | https://static.igem.org/mediawiki/2008/a/a8/Kokodak.jpg" width=300/> <var><b>Fig. 1. Control XbaI/BamHI digests of plasmids isolated from transformants with colony PCR product.</b><br> | ||
| + | 1. Marker<br> | ||
| + | 2-4. digested plasmids pET15b+A-alpha <br></var> | ||
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Revision as of 10:19, 26 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.  Fig. 1. Control XbaI/BamHI digests of plasmids isolated from transformants with colony PCR product.1. Marker 2-4. digested plasmids pET15b+A-alpha
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