Team:Warsaw/Calendar-Main/17 June 2008

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Preparation of constructs with OmpA protein fusions

Michał K.

  1. 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°C).
  2. 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°C).
  3. Gel electrophoresis of PCR products and gel-out of OmpA_alpha (900 bp) and OmpA_omega (700 bp).
  4. Electrophoresis to estimate the concentration of isolated DNA.
  5. Overnight digest of purified OmpA_alpha and OmpA_omega DNA with NdeI and BamHI (Tango 2x buffer).
  6. Inoculation of E. coli TOP10 with pACYC177 plasmid into liquid LB + kanamycin.

Blue/white and rifampicin test

Michał L., Ewa, Marcin

Repetition of experiment from 11/06/2008.