MutD₅ testing

Piotr

Testing MutD₅ ampicillin resistance - MutD₅ has not spontaneous ampicillin resistance mutations.

'Hunter and prey' system tests: Competition tests

Weronika

Inoculation to one tube (0.25 mM IPTG; ampicillin 50 μl/ml)

1. • pACYC177+OmpA_A_omega, pACYC177+OmpA_omega and pACYC177+OmpA_Z_omega - A_alpha protein added ~100 μg
   • pACYC177+OmpA_A_omega, pACYC177+OmpA_omega and pACYC177+OmpA_Z_omega - Z_alpha protein added ~100 μg
   • pACYC177+OmpA_A_alpha, pACYC177+OmpA_alpha and pACYC177+OmpA_Z_alpha - Z_omega protein added ~100 μg

2. Bacteria centrifuged and frozen

Preparation of ΔA (BBa_K103003)

Michał K., Piotr

1. Gel electrophoresis to estimate concentration of purified DNA from previous day.
2. Ligation of isolated DNA fragments of pSB1A3 and ΔA.
3. E. coli TOP10 transformation with the above ligation.
4. Plating on LB + ampicillin.