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Fast cycling with TaqMan detection. An Assays-on-Demand assay (Life Technologies; 171 bp β-actin target) was performed using 50, 5, 0.5 ng human genomic DNA and 10 ng of purified wild-type or mutant Taq DNA polymerase. Reactions were cycled on the StepOnePlus instrument using cycling conditions consisting of 2 min at 95°C followed by 40 cycles of: (A) 10 s at 95°C, 60 s at 60°C; or (B) 10 s at 95°C, 10 s at 60°C.
