Cloning of protein Z DNA to pET15b-OmpA-omega in place of OmpA

Paweł

1. Results of ligation: 6 colonies grown.
2. Each colony cultured overnight in LB + ampicillin.

1. Separate transformant colonies (tranformation of ligation of pACYC177+OmpA_alpha and omega_A from previous day) inoculated to liquid LB with kanamycin.

Antoni:

1. Preparation of chemocompetent bacteria E. coli TOP10.

Cloning omega-A fusion on pKS (second attempt)

Michał L., Ewa, Marcin:

Sequencing confirms that we have obtained proper construct with omega-A fusion

Cloning of protein A DNA to OmpA constructs

1. Colony PCR on colonies from plates with transformations pACYC177+OmpA_A_omega. Primers used: AL+SacI and AP+NotI
2. Confirmed transformant colonies inoculated to liquid LB with kanamycin.
3. Isolation of plasmids from cultures inocluated on previous day (pACYC177+OmpA_A_alpha).
4. Control digest of isolated plasmids with BamHI and SacI.