TABLE 1.

Primer pairs used to amplify MLSA gene sequences, with resulting amplicon lengths, annealing temperatures, and Mg²⁺ concentrations used in PCR^a

MLSA locus	Primer ID	Sequence (5′ to 3′)	Amplicon length (bp)	Anneal temp (°C)	Extra Mg2+ concn (mM)
gyrB	gyrB116b	TGGTTTTTGAGGTGGTGGATA	1,214	60	0.5
	gyrB1330b	CGCTTGATTCTTACGGTTACG	1,214	60	0.5
	gyrB58	CGYAAGCGTCCRGGTATGTA	1,450	55	0.5
	gyrB1507	CGACRTCVGCATCGGTCA	1,450	55	0.5
mdh	mdh540c	GTTTGACGGTCGGATACACC	1,039	60	0.5
	mdh541c	AGAGCGGTATTTTCCAATGC	1,039	60	0.5
	mdh83	CATYGGTCAAGCCCTDGC	678	55	0.5
	mdh760	GCWAGACCRAARCGACAKGC	678	55	0.5
recA	recA884c	TGGACGAGAATAAACAGAAGGC	1,089	60	0.5
	recA885c	AACCTCTTTGCATTCAGCCC	1,089	60	0.5
	recA147	ACCTGAGAGTGACTATCCGG	1,004	55	0.5
	recA1150	GCATTTCACGCAGTTTTTTA	1,004	55	0.5
	recA172	GGYYTACCAATGGGHCGTAT	748	55	0.5
	recA919	TYGCTTTACCTTGRCCRA	748	55	0.5
idh	idh4965b	TTCATTACTGCCGATTATTC	1,063	55	0.5
	idh4966b	TTTGGTGTCTTTCTGCTTAC	1,063	55	0.5
	idh37	ACTGATGAAGCMCCRGCGYT	1,065	55	0.5
	idh1101	DCCCCACATYTGGCCWGA	1,065	55	0.5
asd	asd414d	CCTTTGGCTAAACTCGG	985	50	0.75
	asd416d	GTTATCCGCCACTACCC	985	50	0.75
	asd1051	TGGGATMGHCGYGARTACAGY	811	55	1
	asd1862	RCGKACRCACGTTGGGTT	811	55	1
dnaE	dnaE713c	GATTTCTCTATGGTGGATGG	1,111	60	0.5
	dnaE714c	ATTCCAGCGGATCAAGGTCG	1,111	60	0.5
	dnaE34	CACAGTGATTTYTCDATGGTGG	1,204	55	0.5
	dnaE1237	GRTCACGYTTATCCATACA	1,204	55	0.5
nagB	nagB4	AGACTTATCCCACTGAAAGCG	787	55	0.5
	nagB790	CGATGTTTTTGGCTTCTAACTC	787	55	0.5
	nagB32a	AAGTWGGYAARTGGGCRGC	657	55	0.5
	nagB688	GMAGWGCAGARACKGTCCA	657	55	0.5

^aAll primers were designed for this study unless otherwise noted. Underlining in sequences represents degenerate base positions.

^bPrimer pair published by Salim et al. (46).

^cPrimer pair first published by Byun et al. (5).

^dPrimer pair previously published by Karaolis et al. (27).