Efficiency of cloning in pWXY1.0 under different conditions.

All reaction mixtures contained 1 mM ATP and 5 Weiss unit T4 DNA ligase (Fermentas) and incubated at 37°C. Experiments were performed using 50 ng vector, and the corresponding amount of insert DNA at a 3∶1 molar ratio of insert:vector. None of the reaction mixtures here was inactivated with heat prior to E. coli transformation.
* indicates the reaction mixture was incubated at room temperature 20°C. Cloning efficiencies were given as colonies number on LB plates containing 100 µg/mL ampicillin. Cloning success rate were evulated by colonies PCR test of 50 radom seleted colonies. Specific primers of different inserts used as forward primer, and CYC1 primer in CYC1 terminator used as a reverse primer.
Efficiency of cloning in pWXY1.0 under different conditions.