Bead-based Melting Analysis in Temperature-gradient Microchannels for Single Nucleotide Polymorphisms Detection

A novel single nucleotide polymorphisms (SNPs) detection scheme was proposed by conducting melting analysis on microbeads. These beads were utilized to immobilize target DNA duplex, and flowing through a well-controlled temperature-gradient region in microchannels on a microfluidic chip. As the microbeads going through from low temperature regions to high temperature regions, the DNA duplex denatured and the intercalated fluorescent dyes were released, inducing the fluorescent signals to decay. The melting curves were then acquired for SNP genotyping. Different genotypes (i.e. wild and mutant types) with a SNP location of Ataxia Telangiectasia-Mutated (ATM) genes from Landrace sows were tested and successfully discriminated.