Cloning omega-A fusion on pKS (second attempt)

Michał L., Ewa, Marcin:

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

Checking if omp_omega_A_alpha gives ampicillin resistance
Piotr

1. Inoculation of omp_omega_A_alpha into various IPTG concentrations: 0, 0.1, 0.25, 0.5, 0.75, 1 mmol/mL

Checking the expression of omp_omega_A_alpha and omp_A_alpha
Piotr

1. Inoculation of omp_omega_A_alpha and omp_A_alpha with inductor (0,5 mmol/mL IPTG) and negative control without IPTG.

1. Digest of pACYC177+OmpA_omega_deltaA_alpha (from 25 July) with BamHI and NotI. DNA ends blunting with Klenow fragment (3 hr).
2. Gel elctrophoresis and gel-out of proper band - 4300 bp.
3. Ligation of isolated DNA fragment (1 hr).
4. Transformationof E. coli TOP10 strain with ligation.
5. Transformants plating on LB + kanamycin.