Doitsidou Maria, Jarriault Sophie, Poole Richard J
Centre for Integrative Physiology, University of Edinburgh, EH8 9XD, Scotland
L'Institut de Génétique et de Biologie Moléculaire et Cellulaire, Centre National de la Recherche Scientifique UMR 7104/Institut National de la Santé et de la Recherche Médicale U964, Université de Strasbourg, 67404, France
Genetics. 2016 Oct;204(2):451-474. doi: 10.1534/genetics.115.186197.
The use of next-generation sequencing (NGS) has revolutionized the way phenotypic traits are assigned to genes. In this review, we describe NGS-based methods for mapping a mutation and identifying its molecular identity, with an emphasis on applications in Caenorhabditis elegans In addition to an overview of the general principles and concepts, we discuss the main methods, provide practical and conceptual pointers, and guide the reader in the types of bioinformatics analyses that are required. Owing to the speed and the plummeting costs of NGS-based methods, mapping and cloning a mutation of interest has become straightforward, quick, and relatively easy. Removing this bottleneck previously associated with forward genetic screens has significantly advanced the use of genetics to probe fundamental biological processes in an unbiased manner.
下一代测序（NGS）的应用彻底改变了将表型性状与基因关联的方式。在本综述中，我们描述了基于NGS的用于定位突变并确定其分子身份的方法，重点是在秀丽隐杆线虫中的应用。除了概述一般原理和概念外，我们还讨论了主要方法，提供了实践和概念性的提示，并指导读者进行所需的生物信息学分析类型。由于基于NGS的方法速度快且成本大幅下降，定位和克隆感兴趣的突变变得直接、快速且相对容易。消除以前与正向遗传筛选相关的这一瓶颈极大地推动了以无偏见方式利用遗传学来探究基本生物学过程的发展。
