Team:Warsaw/Calendar-Main/18 July 2008
From 2008.igem.org
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+ | <h3>Cloning of protein A DNA to OmpA constructs</h3> | ||
+ | <p><ol> | ||
+ | <li>Colony PCR on colonies from plates with transformations pACYC177+OmpA_A_omega and pACYC177+OmpA_A_alpha. 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> | <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> | ||
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- | + | <li> Confirmed transformant colonies (found only on pACYC177+OmpA_A_alpha plate) inoculated to liquid LB with kanamycin. </li></ol> | |
</p> | </p> | ||
<p>Paweł: pET15b vector purified. | <p>Paweł: pET15b vector purified. | ||
- | + | Digest of pET15b and pACYClac+OmpA+omega with BamHI and NdeI<br>Gel-out of OmpA+omega (cutted from pACYClac+OmpA+omega, ~1350 bp)</p> | |
<p>Ligation of gel-outed fragment into digested pEt15b | <p>Ligation of gel-outed fragment into digested pEt15b | ||
</p> | </p> | ||
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{{WarNotebookEnd}} | {{WarNotebookEnd}} |
Revision as of 18:21, 1 October 2008
Cloning of protein A DNA to OmpA constructs
Paweł: pET15b vector purified.
Digest of pET15b and pACYClac+OmpA+omega with BamHI and NdeI Ligation of gel-outed fragment into digested pEt15b |