全基因组扩增在人类疾病研究中的应用。

The use of whole genome amplification in the study of human disease.

作者信息

Hughes Simon, Arneson Nona, Done Susan, Squire Jeremy

机构信息

Ontario Cancer Institute, Princess Margaret Hospital, University Health Network, Ontario, Canada.

出版信息

Prog Biophys Mol Biol. 2005 May;88(1):173-89. doi: 10.1016/j.pbiomolbio.2004.01.007.

DOI:10.1016/j.pbiomolbio.2004.01.007

PMID:15561304

Abstract

The availability of large amounts of genomic DNA is of critical importance for many of the molecular biology assays used in the analysis of human disease. However, since the amount of patient tissue available is often limited and as particular foci of interest may consist of only a few hundred cells, the yield of DNA is often insufficient for extensive analysis. To address this problem, several whole genome amplification (WGA) methodologies have been developed. Initial WGA approaches were based on the polymerase chain reaction (PCR). However, recent reports have described the use of non-PCR-based linear amplification protocols for WGA. Using these methods, it is possible to generate microgram quantities of DNA starting with as little as 1mg of genomic DNA. This review will provide an overview of WGA approaches and summarize some of the uses for amplified DNA in various high-throughput genetic applications.

摘要

对于人类疾病分析中使用的许多分子生物学检测而言，获取大量基因组DNA至关重要。然而，由于可获得的患者组织量往往有限，而且特定的感兴趣病灶可能仅由几百个细胞组成，DNA产量常常不足以进行广泛分析。为了解决这个问题，已经开发了几种全基因组扩增（WGA）方法。最初的WGA方法基于聚合酶链反应（PCR）。然而，最近的报告描述了使用基于非PCR的线性扩增方案进行WGA。使用这些方法，从低至1mg基因组DNA开始就有可能生成微克量的DNA。本综述将概述WGA方法，并总结扩增DNA在各种高通量基因应用中的一些用途。