The development of multiple SNPs typing system by flow-arry

DNA序列变异与某种疾病和不同的药物反应有关。单核苷酸多态性（SNP）是常见的变异类型。本研究利用不同大小的微球和一台只有一个激光器的流式细胞仪，开发了一种高通量SNP基因分型的新方法。用Alexa 488标记的聚合酶链反应（PCR）产物与探针杂交，然后通过流式细胞术分析杂交信号。为了对特定的SNP进行分型，将两种珠子与成对的探针结合;一种与野生型序列互补，另一种与变体序列互补。将成对的珠粒放置在同一管中，并且正常珠粒和变体珠粒的数量比例为三比一。当具有较高荧光强度的珠粒是具有较低荧光强度的珠粒的约三倍时，该样品被确定为野生型基因型。当进行多重分析时，每个SNP位点的珠通过其直径来识别。本研究开发了一种新的SNP分型方法，即利用不同大小的微球和配备单激光器的流式细胞仪进行SNP分型。该方法增加了微珠分析的参数

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