The development of multiple SNPs typing system by flow-arry

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DNA sequence variation is associated with a certain disease and differential drug response. Single nucleotide polymorphisms (SNPs) are common type of variant. In this project, we have developed a new method for high-throughput genotyping of SNPs with microspheres of various sizes and a flow cytometer equipped with only a single laserThis system uses microspheres of various sizes with oligonucleotide probes attached by covalent bonds. Polymerase chain reaction (PCR) products labeled with Alexa 488 are hybridized to the probes and then hybridization signals are analyzed by flow cytometry. In order to type a specific SNP, two kinds of beads are bound to paired probes ; one complementary to the wild-type sequence and the other to the variant sequence. The paired beads are placed in the same tube and the number of normal beads and variant beads ratio are three to one. When the beads with higher fluorescence intensity are about three times as many as lower ones, this sample is determined wild-type genotype. When multiplex analysis is carried out, the beads for each SNP site is identified by their diameters. In this project, we have developed a novel method in which SNP genotyping with microspheres of various sizes and flowcytometer equipped with single laser. The method increases parameters in beads analysis