Team:Warsaw/Calendar-Main/17 June 2008

Preparation of constructs: OmpA_alpha and OmpA_omega Michał K.   PCR on OmpA_linker and linker_alpha with

OmpaL_N and AlphaP_XB

primers (20 cycles, elongation length 1 min 15 s, annealing temperature 57&deg;C).  PCR on OmpA_linker and linker_omega with

OmpaL_N and OmegaP_EPB

primers (20 cycles, elongation length 1 min 15 s, annealing temperature 57&deg;C).</li>  Gel electrophoresis of PCR products and gel-out</a> of OmpA_alpha (1000 bp) and OmpA_omega (800 bp). </li> Electrophoresis to estimate the concentration of isolated DNA.</li>  Overnight digest</a> of purified OmpA_alpha and OmpA_omega DNA with NdeI and BamHI (Tango 2x buffer). </li>  Inoculation of E. coli TOP10</a> with pACYC177</a> plasmid into liquid LB + kanamycin. </li> </ol> Blue/white and rifampicin test #2 Michał L., Ewa Repetition of experiment from 11/06/2008.</a>