TABLE 1.
Primers and polymerase chain reaction conditions used in this study.
| Gene | Primer sequence | Size of product (bp) | PCR programme | PCR volume (25 μL) | Reference |
|---|---|---|---|---|---|
| stx1 | F: CTT CGG TAT CCT ATT CCC GG R: GGA TGC ATC TCT GGT CAT TG |
484 | 25 cycles of 30 s at 94 °C 45 s at 50 °C 90 s at 70 °C 10 min at 70 °C |
2.5 μL PCR buffer 10X 1.25 μL MgCl2 0.5 μLdNTP 1 μL of each primers F & R 0.25 μLTaq DNA polymerase 1 μL DNA template |
Tahamtan et al. (2010) |
| stx2 | F: CCA TGA CAA CGG ACA GCA GTT R: CCT GTC AAC TGA GCA GCA CTT TG |
779 | 25 cycles of 30 s at 94 °C 45 s at 50 °C 90 s at 70 °C 10 min at 70 °C |
2.5 μL PCR buffer 10X 1.25 μL MgCl2 0.5 μLdNTP 1 μL of each primers F & R 0.25 μLTaq DNA polymerase 1 μL DNA template |
Tahamtan et al. (2010) |
| eae | F: AAG CGA CTG AGG TCA CT R: ACG CTG CTC ACT AGA TGT |
384 | 25 cycles of 30 s at 94 °C 45 s at 50 °C 90 s at 70 °C 10 min at 70 °C |
2.5 μL PCR buffer 10X 1.25 μL MgCl2 0.5 μLdNTP 1 μL of each primers F & R 0.25 μLTaq DNA polymerase 1 μL DNA template |
Vasconcellos et al. (2012) |
| IntI | F: TGCGGGTYAARGATBTKGATTT* R: CARCACATGCGTRTARAT |
491 | 30 s at 94 °C, 35 s at 57 °C 25 cycles of 1 min at 70 °C 10 min at 72 °C |
2.5 μL PCR buffer X 10 1.25 μL MgCl2 1 μLdNTP 1 μL of each primers F & R 0.25 μLTaq DNA polymerase 1 μl DNA template |
Tahamtan et al. (2014) |
Note: B = C or G or T; K = G or T; R = A or G; Y = C or T*.
PCR, polymerase chain reaction; DNA, deoxyribonucleic acid; Taq, thermus aquaticus; stx, isolates carrying stx1 and/or stx2 genes; eae, isolates carrying eae gene; pb, base pair; IntI, encoded integrases.