Department of Chemistry, University of California, Berkeley, California 94720, United States.
Anal Chem. 2012 Jan 17;84(2):963-70. doi: 10.1021/ac202303f. Epub 2011 Dec 30.
A platform is developed for rapid, multiplexed detection of single-nucleotide polymorphisms using gels copolymerized with oligonucleotide capture probes in a linear microchannel array. DNA samples are analyzed by electrophoresis through the linear array of gels, each containing 20-40 μM of a unique oligonucleotide capture probe. Electrophoresis of target DNA through the capture sites and the high concentration of capture probes within the gels enables significantly shorter incubation times than standard surface DNA microarrays. These factors also result in a significant concentration of target within the gels, enabling precise analysis of as little as 0.6 femtomoles of DNA target. Differential melting of perfectly matched and mismatched targets from capture probes as a function of electric field and temperature enables rapid, unambiguous identification of single-nucleotide polymorphisms.
一个平台被开发出来，用于使用在线性微通道阵列中共聚的寡核苷酸捕获探针快速、多重检测单核苷酸多态性。通过线性阵列的凝胶电泳分析 DNA 样品，每个凝胶中含有 20-40 μM 的独特寡核苷酸捕获探针。目标 DNA 通过捕获位点和凝胶中高浓度的捕获探针电泳，与标准表面 DNA 微阵列相比，孵育时间显著缩短。这些因素还导致目标在凝胶中的浓度显著增加，从而能够精确分析低至 0.6 飞摩尔的 DNA 目标。作为电场和温度函数的捕获探针中完全匹配和错配靶标的差异融解，能够快速、明确地识别单核苷酸多态性。
