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Cycling was performed using an Eppendorf Mastercycler (Hamburg, Germany), as follows: a pre-PCR step of a 5-min denaturation at 94C, followed by 30 cycles of denaturation at 94C for 30 sec, annealing for 30 sec and extension at 72C for 30 sec and a final 5-min incubation at 72C

Prior to that I had abdominal surgery

Ye et al., 2006)

Given an inferior performance of individual biomarkers in diagnosing early PD, combination of multiple biomarkers may greatly improve the diagnostic efficacy