Table 1.

Comparison between Frosst and newly proposed primers’ characteristics

	Frosst et al., 1995		This work
	Forward primer	Reverse primer	Forward primer	Reverse primer
Sequence (5′−3′)	TGAAGGAGAAGGTGTCTGCGGGA	AGGACGGTGCGGTGAGAGTG	TGTGGTCTCTTCATCCCTCGC	CCTTTTGGTGATGCTTGTTGGC
Tm	72°C	66°C	66°C	66°C
Tm difference (optimal <2°C)	6°Ca		0°C
Length (optimal 18−22 nt)	23	20	21	22
GC clamp	Noa	Noa	Yes	Yes
GC content (optimal 40%−60%)	56%	65%a	57%	50%
Secondary structures	No	No	No	No
Overlap between sense primer and Hinf I recognition site.	Yesa	Yesa	No	No
Simple visualization on agarose gel after RFLP	Noa	Noa	Yes	Yes
Presence of other restriction sites for Hinf I	No	No	No	No
Pairing specificity	Yes	Yes	Yes	Yes

^aNot optimal parameter.