<p/>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 µl
template DNA - pKS-A4 1 µl
primer AP+NotI_N - 2 µl
primer AL+link10+homo2_N - 2 µl
Pfu polymerase buffer + Mg2+ (from Fermentas) - 5 µl
dNTPs - 1 µl
Pfu turbo - 0.5 µl
H2o - 38.5 µl

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

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

program: 1. 95°C 3'
2. 95°C 30"
3. 62°C 45"
4. 72°C 45"
5. 72°C 10'
6. keeping in 4°C
25 cycles
3. gel electrophoresis

4.reisolation from agarose gel

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