Table 1.

Marker Positions and PCR-RFLP Conditions for Detection of 13 SNPs in the ACE Gene

	Position
Marker (Location)	Villard et al. (1996)	Present Study	Primer Sequencesa(5′→3′)	TA(°C)	MgCl2(mM)	RFLP Enzyme	T(°C)
ACE1 (5′ UTR)	T−5491C	T−5529C	ACE1F:b TACAACCATCACTACTAATGTCACE1R:b TATAATATATGTGACATGGCCTG	55	2	BstNI	60
ACE2 (5′ UTR)	A−5466C	A−5499C	ACE2F:b GCCATGTCACATATATTATAGGAACE2R:b CGTCTTTGGAAACTTGTCTGC	50	2.5	EcoRV	37
ACE3 (5′ UTR)	T−3892C	T−3925C	ACE3F:b ATAGTGTATATAGGGCTTGGTACACE3R:b AGAAGATATTTGCAAAGTATGTACTG	55	2	PstI	37
ACE4 (5′ UTR)	A−240T	A−262T	ACE4F: TGTCACTCCGGAGGCGGGAGGCTACE4R:b GAGAAAGGGCCTCCTCTCTCT	55	1	XbaI	37
ACE5 (5′ UTR)	C−93T	C−115T	ACE5F2: ACCATGGCCTGGTGAAGAAGCACE5R:b CGGCTCTGCCCCTTCTCCTGCGC	TDc	2.5	HinfI	37
ACEs2.1 (intron 7)		C5144T	ACEs1F: GGGCGGGAAGTGGTGTGCACEs1AR: CTGAAAGCAAGGAAGGAGGAG	62	2	PstI	37
ACEs1.1 (Intron 7)		G5170A	ACEs1CF: GCAGTGAGCTGAGATTGTGCCACEs1AR: CTGAAAGCAAGGAATGAGGAG	62	2	PstI	37
ACE6 (exon 8)	C1237T	C5467T	ACE6F:b AGTGCACACGGGTCACGATGACE6R-1: CACCAAGTAGCCAAAGGGCAG	62	1.5	BsmBI	55
ACENEW 6 (intron 8)		A5967G	ACENEW6F: CAGGGTTCGGGATCCTCCTAGAACE6R-1: CCACCAAGTAGCCAAAGGGCAG	62	1.5	BsmBI	55
ACE7 (exon 15)	A2215G	A9596G	ACE7F:b CACACCCTGAAGTACGGCACACE7R: TCCTCCAGCTCCTGGGCAG	60	1	HaeII	37
ACE I/D (intron 16)			ACEIDF: CTGGAGACCACTCCCATCCTTTCTACEIDR: GATGTGGCCATCACATTCGTCAGAT	62	3
ACE8 (exon 17)	A2350G	A11860G	ACE8F:b CTGACGAATGTGATGGCCGCACE8R:b TTGATGAGTTCCACGTATTTCG	58	2	BstUI	60
ACE9 (3′ UTR)	4656 (TC)2/3	21288 (CT)2/3	ACE9FB: GACTCTGGGAAGCAGACATACE9RA: AGCTAATCCCTGTGCAGGT	(See text)	(See text)	(See text)	(See text)

^aUnderlined nucleotides represent the deliberate primer mismatches, designed to introduce artificial restriction sites.

^bDescribed by Keavney et al. (1998).

^cTouchdown PCR.