TABLE 2.

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

GenePrimersaReference or sourceSize (nt)PCR thermocycling parameters
cpn60 GAI III GCI GGI GAY GGI ACI ACI AC 12 60094°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 GTCNCBIc90694°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 80095°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 GNCBId97694°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 48095°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 80395°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
  • a All 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).

  • b Touchdown PCR from 45°C to 35°C with temperature decrease of 0.5°C per cycle.

  • c Accession numbers AE006494 , AE009973 , M95569 , X97788 , and Y12602 .

  • d Accession numbers AJ006413 , AJ012548 , AJ012549 , AJ131604 , AJ131605 , and AJ131631 .