Spiked GBS: a unified,open platform for single marker genotyping and whole-genome profiling |
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Authors: | Trevor W Rife Shuangye Wu Robert L Bowden Jesse A Poland |
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Affiliation: | .Interdepartmental Genetics, Kansas State University, 4024 Throckmorton Hall, Manhattan, KS 66506 USA ;.Wheat Genetics Resource Center, Department of Plant Pathology, Kansas State University, 4024 Throckmorton Hall, Manhattan, KS 66506 USA ;.USDA-ARS, Hard Winter Wheat Genetics Research Unit, 4008 Throckmorton Hall, Manhattan, KS 66506 USA ;.Department of Plant Pathology, Kansas State University, Manhattan, KS 66506 USA |
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Abstract: | BackgroundIn plant breeding, there are two primary applications for DNA markers in selection: 1) selection of known genes using a single marker assay (marker-assisted selection; MAS); and 2) whole-genome profiling and prediction (genomic selection; GS). Typically, marker platforms have addressed only one of these objectives.ResultsWe have developed spiked genotyping-by-sequencing (sGBS), which combines targeted amplicon sequencing with reduced representation genotyping-by-sequencing. To minimize the cost of targeted assays, we utilize a small percent of sequencing capacity available in runs of GBS libraries to “spike” amplified targets of a priori alleles tagged with a different set of unique barcodes. This open platform allows multiple, single-target loci to be assayed while simultaneously generating a whole-genome profile. This dual-genotyping approach allows different sets of samples to be evaluated for single markers or whole genome-profiling. Here, we report the application of sGBS on a winter wheat panel that was screened for converted KASP markers and newly-designed markers targeting known polymorphisms in the leaf rust resistance gene Lr34.ConclusionsThe flexibility and low-cost of sGBS will enable a range of applications across genetics research. Specifically in breeding applications, the sGBS approach will allow breeders to obtain a whole-genome profile of important individuals while simultaneously targeting specific genes for a range of selection strategies across the breeding program.Electronic supplementary materialThe online version of this article (doi:10.1186/s12864-015-1404-9) contains supplementary material, which is available to authorized users. |
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Keywords: | Plant breeding Wheat Marker platform Genotyping-by-sequencing |
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