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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.