Microfluidic Genotyping by Rapid Serial PCR and High-Speed Melting Analysis

samples to minimize costs or uses multiplex lab-on-a-chip disposables to analyze a few targets. In genetics,mul-tiple variants need to be analyzed, and different work flows that rapidly analyzemultiple loci in a few targets are attractive. METHODS: We used a microfluidic platform tailored to rapid serial PCR and high-speedmelting (HSM) to geno-type 4 single nucleotide variants. A contiguous stream of master mix with sample DNA was pulsed with each primer pair for serial PCR and melting. Two study sites each analyzed 100 samples for F2 (c.*97GA), F5 (c.1601GA), and MTHFR (c.665CT and c.1286AC) after blinding for genotype and genotype proportions. Internal temperature controls improved melting curve precision. The platform’s liquid-handling system auto