Ni Fei, Yu Yang, Epstein Lynn, Fu Daolin, Wu Jiajie
State Key Laboratory of Wheat Improvement, College of Agronomy, Shandong Agricultural University, Tai'an, Shandong, China.
Department of Plant Pathology, University of California, Davis, CA, USA.
Methods Mol Biol. 2025;2898:291-305. doi: 10.1007/978-1-0716-4378-5_19.
Sequencing trait-associated mutations (STAM) is a simple and straightforward gene cloning method that was developed in wheat. It uses full-length isoform sequencing (Iso-Seq) of the wild type as the reference and employs transcriptome sequencing of multiple, independently derived mutants for gene cloning. The STAM method eliminates the need for fine-mapping or a high-quality whole genome assembly of a specific wheat cultivar, and it could also be used in other plant species with complex genomes. Detailed, bioinformatic analysis protocol and tips for STAM are provided in this chapter.
测序性状关联突变(STAM)是一种在小麦中开发的简单直接的基因克隆方法。它以野生型的全长异构体测序(Iso-Seq)作为参考,并利用多个独立衍生突变体的转录组测序进行基因克隆。STAM方法无需对特定小麦品种进行精细定位或高质量的全基因组组装,也可用于其他具有复杂基因组的植物物种。本章提供了STAM详细的生物信息学分析方案和提示。