微流控液滴单分子PCR.doc

 n  微流控液滴单分子 PCR# 朱志，王春明，官志超，杨朝勇** （厦门大学化学化工学院化学生物学系，固体表面物理化学国家重点实验室，分析科学重点 5 10 实验室，福建 厦门 361005） 摘要：近年来，微流控液滴 PCR 技术被广泛地应用于单分子的放大及分析。微流控液滴技术 具有许多优异的特性，如能将反应分割在多个独立的空间内进行，在单分散的液滴中同时进 行多个平行反应，避免了液滴间的交叉污染，消除 PCR 偏好与非特异性放大，以及进行快速 的热循环从而实现高效放大。本文将对近五年来微流控液滴领域的重大技术突破以及它们在 单分子放大分析中的应用如高通量筛选、下一代 DNA 测序技术、稀有突变检测等进行评述。 虽然目前微流控液滴单分子 PCR 目前还在起步阶段，但其已展现了巨大的应用前景，未来有 望继续为生物医药工程领域在单分子放大分析面临的挑战提供新的解决思路。 关键词：单分子放大；乳液 PCR；微流控液滴；高通量；稀有突变检测 中图分类号：O652.6 15 Microfluidic Droplets for Single-Molecule PCR ZHU Zhi, WANG Chunming, GUAN Zhichao, YANG James Chaoyong (State Key Laboratory of Physical Chemistry of Solid Surfaces, Department of Chemical Biology, Key Laboratory of Analytical Science, College of Chemistry and Chemical Engineering, Xiamen 20 25 30 University, FuJian XiaMen 361005) Abstract: The application of microfluidic droplet PCR for single-molecule amplification and analysis has recently been extensively reported. Microfluidic droplet technology has the advantages of compartmentalizing reactions into discrete volumes, performing highly parallel reactions in monodisperse droplets, reducing cross-contamination between droplets, eliminating PCR bias and nonspecific amplification, as well as enabling fast amplification with rapid thermocycling. Here, we have reviewed the important technical breakthroughs of microfluidic droplet PCR in the past five years and their application to single-molecule amplification and analysis, such as high-throughput screening, next generation DNA sequencing, and quantitative detection of rare mutations. Although the utilization of microfluidic droplet single-molecule PCR is still in the early stages, its great potential has already been demonstrated and will provide novel solutions to today’s biomedical engineering challenges in single-molecule amplification and analysis. Keywords: Single-molecule amplification; emulsion PCR; microfluidic droplets; high-throughput; rare mutation detection 35 0 前言 有选择性地针对单分子尤其是单个生物分子的灵敏检测对化学、生物、医疗以及环境科 学都有着重大而