Preparation of constructs with OmpA protein fusions
1. Colony PCR on colonies from plates with transformations OmpA_alpha.
2. Confirmed transformant colonies inoculated to liquid LB with kanamycin.
Cloning of protein Z DNA to OmpA constructs
1. 2 colonies was inoculated to liquid LB broth with kanamycin

Preparation of construct omega-A

1. PCR A in 50 ul
template DNA - pKS-A4 1ul
primer APNot - 2 ul
primer ALhomo2 - 2 ul
Pfu polymerase buffer + Mg2+ (from Fermentas) - 5 ul
dNTPs - 1 ul
Pfu turbo - 0.5 ul
H2o - 38.5 ul

program: 1. 95oC 3'
2. 95oC 30"
3.62oC 45"
4.72oC 45"
5.72oC 10'
6. keeping in 4oC

2. PCR omega in 50 ul
template DNA - pUC19 1ul
primer OmegaLS - 2 ul
primer AOmegaPli - 2 ul
Pfu polymerase buffer + Mg2+ (from Fermentas) - 5 ul
dNTPs - 1 ul
Pfu turbo - 0.5 ul
H2o - 38.5 ul

program: 1. 95oC 3'
2. 95oC 30"
3. 62oC 45"
4. 72oC 45"
5. 72oC 10'
6. keeping in 4oC
25 cycles </p>

3. gel electrophoresis

4.reisolation from agarose gel