Table 2.

Deletion mutagenesis efficiency results for the different protocols used. Five clones were analysed for each of the different PCR-based mutagenesis methods based on duplicate PCR experiments.

Primer pair	Mutagenesis method	PCR product analysed with agarose gel electrophoresis	Restriction enzyme mapping with HindIII	Deletion efficiency confirmed with nucleotide sequencing (%)
P1	QuickChange™ site-directed method	No product	NA	0
P2	Overlapping primer method	7 kb	~3900 bp ~3100 bp	40
P3	ExSite™ method	No product	NA	0
	Inverse PCR method	7 kb	~3900 bp 3100 bp	0
	RE-mediated inverse PCR method	7 kb	~3900 bp ~3100 bp	80
P4	RE-mediated inverse PCR method	4.6 kb	EcoRI linearization site removed	100