From 2008.igem.org
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| - | <h3>Cloning of protein A DNA to OmpA constructs</h3>
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| - | <ol><li>Colony PCR on colonies from plates with transformations pACYC177+OmpA_A_omega. Primers used:
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| - | <a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#AL+SacI">AL+SacI</a> and <a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#AP+NotI">AP+NotI</a></li>
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| - | <li>Confirmed transformant colonies inoculated to liquid LB with kanamycin. </li>
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| - | <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 (pACYC177+OmpA_A_alpha). </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 with BamHI and SacI.</li></ol>
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| - | </p>
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Revision as of 20:23, 11 October 2008
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Cloning of protein Z DNA to pET15b-OmpA-omega in place of OmpAPaweł
- Results of ligation: 6 colonies grown.
- Each colony cultured overnight in LB + ampicillin.
- Separate transformant colonies (tranformation of ligation of pACYC177+OmpA_alpha and omega_A from previous day) inoculated to liquid LB with kanamycin.
Antoni:
- Preparation of chemocompetent bacteria E. coli TOP10.
Cloning omega-A fusion on pKS (second attempt)
Michał L., Ewa, Marcin:
Sequencing confirms that we have obtained proper construct with omega-A fusion
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