Five μL of the ligated vector was cloned into 200 μL of competent Escherichia coli (DH5?) cells by heat treatment at 42 °C for 45 s and the whole content was transferred into a tube containing
800 μL of SOC (tryptone - 2% w/v, yeast extract -0.5% w/v, NaCl - 8.6 mM, KCl - 2.5 mM, MgSO4 - 2.0 mM, glucose - 20 mM in 1000 mL water, pH 7.0) and rotated at 150 rpm, 37 °C for 1 h; 200 μL
of the above culture was spread on Luria Bertani
agar (LBA) (tryptone - 10 g, yeast extract - 5 g,