Team:Warsaw/Calendar-Main/21 July 2008

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<h3>Cloning of protein A DNA to pET15b+Omp-alfa plasmid<br>Antoni</h3>
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<li> Transformation of <i>E. coli</i> <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top">TOP10</a> strain with friday ligation. </li>
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<li> Transformants plating on LB + ampitilin.</li>
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Revision as of 23:33, 2 October 2008

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Cloning of protein A DNA to OmpA constructs
1. Colony PCR on colonies from plates with transformations pACYC177+OmpA_A_omega. Primers used: AL+SacI and AP+NotI
2. Confirmed transformant colonies inoculated to liquid LB with kanamycin.
3. Isolation of plasmids from cultures inocluated on previous day (pACYC177+OmpA_A_alpha).
4. Control digest of isolated plasmids with BamHI and SacI .

Cloning of protein Z DNA to pET15b-OmpA-omega in place of OmpA
Paweł

  1. Ligations transformed into TOP10 and plated on LB + ampicillin

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