Preparation of constructs with OmpA protein fusions
Michał K.
Inoculation other separate transformant colonies from plates with ligations: pACYC177+OmpA_alpha pACYC177+OmpA_omega (liquid LB + kanamycin).
Preparation of alpa-A and omega-A fusions
Michał L., Ewa, Marcin
We have successfully amplified Omega-linker (Fig. 1.), but were unable to obtain the other two. PCRs were repeated. New annealing temp.: 48°C. Mg2+ concentration elevated to 4 mM.
Fig. 1.PCR products. Lane 1 - lack of product for A-linker; lane 3 - product for Omega-linker.