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Table 4 PCR primers and conditions for multilocus sequence analysis

From: Analysis of Escherichia coli O157 clinical isolates by multilocus sequence typing

Gene

Primer sequencea

Reaction parametersb

Amplicon size (bp)

Gene ID

Locus tag

arcA

F: 5'-GAAGACGAGTTGGTAACACG-3'

R: 5'-CTTCCAGATCACCGCAGAAGC-3'

95°C for 1 min, 55°C for 2 min, 72°C for 3 min, 30 cycles

680

959654

Z6004

aroE

F: 5'-AAGGTGCGAATGTGACGGTG-3'

R: 5'-AACTGGTTCTACGTCAGGCA-3'

95°C for 1 min, 57°C for 2 min, 72°C for 3 min, 28 cycles

620

961663

Z2720

dnaE

F: 5'-GAG/TATGTGTGAGCTGTTTGC-3'

R: 5'-CGA/GATA/CACCGCTTTCGCCG-3'

94°C for 45 s, 45°C for 45 s, 72°C for 1 min, 30 cycles

550

956917

Z0196

mdh

F: 5'-CAACTGCCTTCAGGTTCAGAA-3'

R: 5'-GCGTTCTGGATGCGTTTGGT-3'

94°C for 45 s, 50°C for 45 s, 72°C for 1 min, 30 cycles

580

958670

Z4595

gnd

F: 5'-GGCTTTAACTTCATCGGTAC-3'

R: 5'-TCGCCGTAGTTCAGATCCCA-3'

94°C for 45 s, 50°C for 45 s, 72°C for 1 min 10 s, 30 cycles

590

962087

Z3191

gapA

F: 5'-GATTACATGGCATACATGCTG-3'

R: 5'-CAGACGAACGGTCAGGTCAAC-3'

94°C for 45 s, 50°C for 45 s, 72°C for 1 min 10 s, 30 cycles

535

961753

Z2818

pgm

F: 5'-CCGTCCCATAACCCGCCGGAAG

ATGGTGGTATCAAGTACAATCCG-3'

R: 5'-TTACGCGTTTTTCAGAACTTCGCT

AACAATCTCAACCGCTTCTTT-3'

94°C for 45 s, 50°C for 45 s, 72°C for 1 min, 35 cycles

1146

957767

Z0837

espA

F: 5'-ATGGATACATCAAATGCAACATC

CGTTGTTAATGTGAGTGCGAGT-3'

R: 5'-TTATTTACCAAGGGATATTGCTG

AAATAGTTCTATATTGTAGAGA-3'

94°C for 45 s, 50°C for 45 s, 72°C for 1 min, 35 cycles

579

960865

Z5107

ompA

F: 5'-AGACAGCTATCGCGATTGC-3'

R: 5'-GCTTTGTTGAAGTTGAACAC-3'

94°C for 45 s, 50°C for 45 s, 72°C for 1 min, 30 cycles

691

958948

Z1307

  1. aF, forward; R, reverse.
  2. bAll reactions had an initial denaturation at 94°C for 4 min and a final extension at 72°C for 4 min.