Figure 4

Electrophoretic analyses of multiplex PCR amplicons: etpC (219 pb); stx1 (302 bp); stx2 (519 bp); and eae (346 bp). A. Temperature gradient of annealing step in the multiplex PCR using E. coli EDL933 as DNA template. Line 1 presents the anealing temperature of 55°C; line 2: 55.3°C; line 3: 55.8°C; line 4: 56.7°C; line 5: 57.8°C; line 6: 59.3°C; line 7: 61°C; line 8: 62.4°C; line 9: 63.5°C; line 10: 64.3°C; and line 11: 65°C. For all and 3-minute elongation step was performed.B. Genomic DNAs from E. coli strains: (1) EDL933, (2) CB571, (3) 86–24, (4) DH5α, (5) MG1655, and (6) W3110, were applied as the PCR templates, respectively. Reaction conditions: 2-minute primer annealing at 56°C, and 3-minute elongation step. NC is a blank PCR assay. L stands for DNA Ladder.