Team:Warsaw/Calendar-Main/12 July 2008
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1. Isolation of plasmids from cultures inocluated on previous day (pCACYC177 + OmpA_alpha and pACYC177+OmpA_Z_omega). <br> | 1. Isolation of plasmids from cultures inocluated on previous day (pCACYC177 + OmpA_alpha and pACYC177+OmpA_Z_omega). <br> | ||
2. Control digestation of isolated plasmids with BamHI and SacI (we found good clones for both ligations).</p> | 2. Control digestation of isolated plasmids with BamHI and SacI (we found good clones for both ligations).</p> | ||
+ | |||
+ | <p>'''Linked PCR Omega-A'''<br> | ||
+ | |||
+ | reisolated PCR product PCR-omega - 4 µl<br> | ||
+ | reisolated PCRT product A-homo - 13.5 µl<br> | ||
+ | primer OmegaLS - 2 µl<br> | ||
+ | primer APNot - 2 µl<br> | ||
+ | Pfu buffer with Mg2+ - 5 µl<br> | ||
+ | dNTPs - 1 µl<br> | ||
+ | H2o - 22 µl<br> | ||
+ | |||
+ | program:<br> | ||
+ | |||
+ | 1. 95°C - 3' | ||
+ | 2. 95°C - 30" | ||
+ | 3. 55°C - 45" | ||
+ | 4. 68°C - 1' | ||
+ | 5. go to step 2 25 x | ||
+ | 6. 68° - 10' | ||
+ | 7. keep in 4° | ||
+ | |||
+ | gel electrophoresis<br> | ||
Revision as of 16:48, 26 September 2008