Table 1. Selected RE recognition sites with compatible cohesive ends.

				NEB Buffer activity (%)
Usage	Enzyme	Site	Compatible	1	2	3	4	PCR
c1 MCS	Nhe I	G|CTAGC	Spe I / Xba I	100	100	10	100	100
c1 MCS, c1 sub	Bsr GI	T|GTACA	Bsi WI	25	100	10	100	<25
c1 PCR, c1 sub	Spe I	A|CTAGT	Nhe I / Xba I	75	100	25	75	100
c1 PCR	* Bsi WI	C|GTACG	Bsr GI	100	100	100	25	50
c2 MCS	Asc I	GG|CGCGCC	Mlu I	0	10	10	100	100
c2 MCS, c2 sub	Pac I	TTAAT|TAA	Asi SI	100	75	10	100	100
c2 PCR, c2 sub	Mlu I	A|CGGCT	Asc I	25	75	100	50	50
c2 PCR	Asi SI	GCGAT|CGC	Pac I	50	100	100	50	100

Four pairs of RE recognition sites with compatible cohesive ends were suitable for the plasmid used in this study. The 4 pairs were divided into 2 ‘core’ sets so that the enzymes with the most similar buffer requirements were grouped together, based on the % activity of each enzyme in the 4 different New England Biolabs buffers plus standard PCR buffer (catalog & Technical Reference, 2007–08). ^* Bsi WI was optimally active at 55°C and 50 % active at 37°C.