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Table 1 Details of the PCR protocols tested. All protocols included substitution of dTTP with dUTP and commenced with a uracil N-glycosylase (UNG) incubation step (37°C for 30 minutes) followed by a HotStarTaq DNA Polymerase (Qiagen) activation/UNG inactivation step (95°C, 15 min).

From: Touchdown General Primer (GP5+/GP6+) PCR and optimized sample DNA concentration support the sensitive detection of human papillomavirus

Protocol Denaturation Touchdown annealing cycles Additional annealing cycles Extension Final extension
GP5+/GP6+ 1 min, 94°C - 2 min, 40°C (40–50 cycles) 1.5 min, 72°C 4 min, 72°C
TDP1 1 min, 94°C 2 min, 45°C to 40°C in 0.5°C decrements (11 cycles) 2 min, 40°C (29–39 cycles) 1.5 min, 72°C 4 min, 72°C
TDP2 1 min, 94°C 2 min, 50°C to 40°C in 1.0°C decrements (11 cycles) 2 min, 40°C (29–39 cycles) 1.5 min, 72°C 4 min, 72°C
TDP3 1 min, 94°C 2 min, 50°C to 40°C in 0.5°C decrements (21 cycles) 2 min, 40°C (19–29 cycles) 1.5 min, 72°C 4 min, 72°C
TDP4 1 min, 94°C 2 min, 55°C to 40°C in 1.0°C decrements (16 cycles) 2 min, 40°C (24–34 cycles) 1.5 min, 72°C 4 min, 72°C
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BMC Clinical Pathology

ISSN: 1472-6890

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