Tsai Helen, Ngo Kathie, Lieberman Meric, Missirian Victor, Comai Luca
Department of Plant Biology and Genome Center, University of California at Davis, One Shields Avenue, Davis, CA, 95616, USA.
Methods Mol Biol. 2015;1284:359-80. doi: 10.1007/978-1-4939-2444-8_18.
TILLING is a method to find mutations in a gene of interest by scanning amplicons from a mutagenized population for sequence changes, commonly a single nucleotide. In the past 5 years, mutation detection by sequencing has become increasingly popular. This chapter details the experimental flow for TILLING-by-Sequencing, highlighting the critical steps involved in tridimensional pooling of genomic DNA templates, preparation of libraries for high-throughput sequencing, and bioinformatic processing of the sequence data.
定向诱导基因组局部突变（TILLING）是一种通过扫描诱变群体的扩增子以寻找目标基因中的突变（通常是单核苷酸突变）的方法。在过去5年中，通过测序进行突变检测越来越普遍。本章详细介绍了基于测序的TILLING实验流程，重点强调了基因组DNA模板三维混合、高通量测序文库制备以及序列数据的生物信息学处理等关键步骤。
