Abstract: Restriction-site-associated DNA sequencing (RAD-seq) is a technology of reduced-representation genome sequencing (RRGS), which is developed with next-generation sequencing. In terms of RAD-seq, genome is digested by restriction endonuclease. The digested fragments are ligated with barcoded adapters, and used to construct library for sequencing. With the advantages of simple operation, low experimental cost, and high throughput, RAD-seq is widely used in molecular ecology, evolutionary genomics and conservation genetics. Recently, five different RAD-seq methods including mbRAD, ddRAD, 2bRAD, ezRAD and nextRAD have been developed. Of them, mbRAD uses a low frequency cutter and physical shearing to make genome into fragments. ddRAD uses a low frequency cutter and a high frequency cutter to digest genome. 2bRAD relies on IIB-type restriction endonuclease to excise genomic sequences into short fragments. ezRAD uses isochimozer enzymes and Illumina library preparation kit. nextRAD entirely uses Nextera DNA library preparation kit. In this article, we reviewed the characteristics of each RAD-seq method and its application in entomology, such as population genetics, community ecology, species delimitation, phylogenetic reconstruction and construction of genetic linkage map.    

Key words:  RAD-seq, entomology, population genetics, species delimitation, phylogeny, genetic linkage map