Table 1. Assembly of pGFPi from 10 plasmids.

Ligation conditions	neg	3 h 37°C	6 h 37°C		25 cycles		50 cycles
	blue/white	20 µl plated blue/white	20 µl plated blue/white	estimated total correct	20 µl plated blue/white	estimated total correct	20 µl plated blue/white	estimated total correct
Normal Ligase	1517/0	3/1	1/13	568	1/131	5731	1/379	16581
HC Ligase	985/0	0/72	0/181	7918	1/256	11200	0/211	9231

Restriction-ligation was performed with continuous incubation at 37°C or for 25 or 50 cycles (2 min 37°C+5 min 16°C). 20 µl out of 1 ml were plated. The negative control was performed without BsaI enzyme.