a robust, simple genotyping-by-sequencing (gbs) approach for high diversity species一个健壮的、简单的genotyping-by-sequencing(gbs)方法对高多样性的物种.pdfVIP

A Robust, Simple Genotyping-by-Sequencing (GBS) Approach for High Diversity Species 1 1 2 3 1 1,4 Robert J. Elshire , Jeffrey C. Glaubitz , Qi Sun , Jesse A. Poland , Ken Kawamoto , Edward S. Buckler , Sharon E. Mitchell1* 1 Institute for Genomic Diversity, Cornell University, Ithaca, New York, United States of America, 2 Computational Biology Service Unit, Cornell University, Ithaca, New York, United States of America, 3 Hard Winter Wheat Genetics Research Unit, United States Department of Agriculture/Agricultural Research Service, Manhattan, Kansas, United States of America, 4 Plant, Soil and Nutrition Research Unit, United States Department of Agriculture/Agricultural Research Service, Ithaca, New York, United States of America Abstract Advances in next generation technologies have driven the costs of DNA sequencing down to the point that genotyping-by- sequencing (GBS) is now feasible for high diversity, large genome species. Here, we report a procedure for constructing GBS libraries based on reducing genome complexity with restriction enzymes (REs). This approach is simple, quick, extremely specific, highly reproducible, and may reach important regions of the genome that are inaccessible to sequence capture approaches. By using methylation-sensitive REs, repetitive regions of genomes can be avoided and lower copy regions targeted with two to three fold higher efficiency. This tremendously simplifies computationally challenging alignment problems in species with high levels of genetic diversity. The GBS procedure is demonstrated with maize (IBM) and barley (Oregon Wolfe Barley) recombinant inbred populations where roughly 200,000 and 25,000 sequence tags were mapped,