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Team:Warsaw/Calendar-Main/2 July 2008
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Preparation of constructs with OmpA protein fusions
PCR on OmpA_linker and linker_alpha with
OmpaL_N
and
AlphaP_XB
primers (20 cycles, annealing length 1 min 15 s, annealing temperature 57°C)
PCR on OmpA_linker and linker_omega with
OmpaL_N
and
OmegaP_EPB
primers (20 cycles, annealing length 1 min 15 s, annealing temperature 57°C)
Gel electrophoresis of PCR products and
gel-out
of OmpA_alpha and OmpA_omega
Overnight
digest
of purified OmpA_alpha and OmpA_omega DNA with NdeI and BamHI
Overnight
digest
of pET15b plasmid with NdeI and BamHI,
dephosphorylation with CIAP