Team:Warsaw/Calendar-Main/21 July 2008

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4. Control digest of isolated plasmids with BamHI and SacI .
4. Control digest of isolated plasmids with BamHI and SacI .
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<p>Paweł: restriction analysis of plasmids isolated from overnight cultures. Result: none of isolated plasmids contain insert</p>
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<h3>Cloning of protein Z DNA to pET15b-OmpA-omega in place of OmpA<br>
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<p>Another ligation prepared</p>
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Paweł</h3>
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<p><ol><li>Ligations transformed into TOP10 and plated on LB + kanamycin</li></ol></p>

Revision as of 16:50, 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 + kanamycin

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