TABLE 2.

Primer sequences and source information, amplicon size, and thermocycling parameters for MLST of S. uberis

Gene	Primersa	Reference or source	Size (nt)	PCR thermocycling parameters
cpn60	GAI III GCI GGI GAY GGI ACI ACI AC	12	600	94°C, 2:00; 20× (94°C, 1:00; TDb, 2:00; 72°C, 5:00); 20× (94°C, 1:00; 35°C, 2:00; 72°C, 5:00); 72°C, 10:00
	YKI YKI TCI CCR AAI CCI GGI GCY TT
gapC	TTG GTA TTA ACG GTT TCG GTC	NCBIc	906	94°C, 9:00; 35× (94°C, 1:00; 50°C, 1:00; 72°C, 1:00); 72°C, 5:00
	CAA GTT GAG CAG TGT AAG ACA TTT C
oppF	GAA GCG AAG CTT TGG CT GG	43	800	95°C, 4:00; 35× (95°C, 1:00; 55°C, 1:00; 72°C, 1:00); 72°C, 7:00
	GCA GCT TCT GCT TCT GTT GA
pauA	TTC ACT GCT GTT ACA TAA CTT TGT G	NCBId	976	94°C, 5:00; 35× (94°C, 1:00; 50°C, 1:00; 72°C, 1:00); 72°C, 5:00
	CCT TTG AAA GTG ATG CTC GTG
sodA	CCI TAY ICI TAY GAY GCI YTI GAR CC	40	480	95°C, 3:00; 35× (95°C, 0:30; 37°C, 2:00; 72°C, 1:30); 72°C, 10:00
	ARR TAR TAI GCR TGY TCC CAI ACR TC
tuf	AAY ATG ATI ACI GGI GCI CAR ATG GA	24	803	95°C, 3:00; 35× (95°C, 0:30; 55°C, 0:30; 72°C, 1:00); 72°C, 7:00
	AYR TTI TCI CCI GGC ATI ACC AT

^aAll primers are shown 5′ to 3′. For each gene, the first primer listed is the forward primer, and the second primer is the reverse primer. I = inosine; K = keto (G or T); R = purine (A or G); Y = pyrimidine (C or T).

^bTouchdown PCR from 45°C to 35°C with temperature decrease of 0.5°C per cycle.

^cAccession numbers AE006494, AE009973, M95569, X97788, and Y12602.

^dAccession numbers AJ006413, AJ012548, AJ012549, AJ131604, AJ131605, and AJ131631.