Bilichak Andriy, Golubov Andrey, Kovalchuk Igor
Lethbridge Research Centre, Agriculture and Agri-Food Canada, Lethbridge, AB, Canada.
Department of Biological Sciences, University of Lethbridge, 4401 University Drive, Lethbridge, AB, Canada, T1K 3M4.
Methods Mol Biol. 2017;1456:189-196. doi: 10.1007/978-1-4899-7708-3_15.
The discovery of small RNAs in plants and animals almost two decades ago attracted a significant interest towards epigenetic regulation of gene expression and the practical implementation of the gained knowledge in applied studies. New and sometimes unexpected functions have been ascribed to sRNAs almost every couple of years since their discovery, hence indicating that the complete role of sRNAs in plant and animal physiology is still barely understood. Next-generation sequencing technologies allow to generate high-resolution profiles of sRNAs for the consequent analysis and possibly to discover novel functions of sRNAs. In this chapter, we provide brief guidelines for sRNA library preparation in plants and a practical approach that can be implemented to overcome possible difficulties with sequencing library generation.
大约二十年前，植物和动物中小RNA的发现引发了人们对基因表达表观遗传调控以及将所获知识应用于实际研究的浓厚兴趣。自发现以来，几乎每隔几年就会赋予小RNA新的、有时甚至意想不到的功能，这表明小RNA在植物和动物生理学中的完整作用仍鲜为人知。新一代测序技术能够生成小RNA的高分辨率图谱，以便进行后续分析，并有可能发现小RNA的新功能。在本章中，我们提供了植物小RNA文库制备的简要指南，以及一种可用于克服测序文库构建可能遇到的困难的实用方法。
