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Table 1 Oligonucleotide primers used for polymerase chain reaction (PCR), PCR conditions, and product size of vacA alleles and cagA

From: The clearance effect of bovine anti-Helicobacter pylori antibody-containing milk in O blood group Helicobacter pylori-infected patients: a randomized double-blind clinical trial

Genes Primers Product size (bp) Initial denaturation temperature °C (min) Denaturation temperature °C (min) Annealing temperature °C (min) Extension temperature °C (min) Cycles Final extension temperature °C (min)
s1a F:5′-GTCAGCATCACACCGCAAC-3′ 190   94 (1) 52 (1) 72 (1) 35 72 (5)
R:5′-CTGCTTGAATGCGCCAAAC-3′
s1b F:5′-AGCGCCATACCGCAAGAG-3′ 187   94 (1) 52 (1) 72 (1) 35 72 (5)
R:5′-CTGCTTGAATGCGCCAAAC-3′
s2 F: 5′-GCTAACACGCCAAATGATCC-3′ 199   94 (1) 52 (1) 72 (1) 35 72 (5)
R: 5′-CTGCTTGAATGCGCCAAAC-3′
m1a F: 5′-GGTCAAAATGCGGTCATGG-3′ 290 95 (2) 94 (0.5) 52 (0.5) 72 (0.5) 40 72 (5)
R: 5′-CCATTGGTACCTGTAGAAAC-3′
m1b F: 5′-GGCCCCAATGCAGTCATGGAT-3′ 240–270 95 (2) 94 (0.5) 52 (0.5) 72 (0.5) 40 72 (5)
R: 5′-GCTGTTAGTGCCTAAAGAAGCAT-3′
m2 F: 5′-GGAGCCCCAGGAAACATTG-3′ 352 95 (2) 94 (0.5) 52 (0.5) 72 (0.5) 40 72 (5)
R: 5′-CATAACTAGCGCCTTGCAC-3′
cagA F:5′-TTGACCAACAACCACAAACCGAAG-3′ 183 94 (2) 95 (0.5) 50 (0.75) 72 (0.75) 40 72 (5)
R:5′-CTTCCCTTAATTGCGAGATTCC-3′
  1. Detection of vacA alleles and cagA. The vacA and cagA genotyping was performed by PCR by using specific primers, as described in Table 1. PCR was conducted in 25-μL volumes containing 1 × PCR buffer, 1 μL of the DNA template, 0.2 mmol/L dNTP mixes, 6 pmol of each primer, and 0.75 U Taq polymerase (New England Biolabs, Ipswitch, MA, USA). The amplified products were electrophoresed on a 1.5% agarose gel, stained with ethidium bromide, and visualized under ultraviolet illumination.