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.

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

1. Ligations transformed into TOP10 and plated on LB + ampicillin.

Cloning omega-A fusion on pKS (second attempt)

Michał L., Ewa, Marcin:

1. We have obtained white colonies of transformants and inoculated 10 of them on liquid LB+Amp₁₀₀

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

1. Result of transformation of pET15b+omega with protein Z DNA: 4 colonies grown
2. Each colony cultured overnight in LB+amp

Michał L., Ewa, Marcin

We have finally got rid of phage infection and we are able to work with E. coli again. Hurray!