PT - JOURNAL ARTICLE AU - Alina Ott AU - Sanzhen Liu AU - James C. Schnable AU - Cheng-Ting “Eddy” Yeh AU - Cassy Wang AU - Patrick S. Schnable TI - Tunable Genotyping-By-Sequencing (tGBS®) Enables Reliable Genotyping of Heterozygous Loci AID - 10.1101/100461 DP - 2017 Jan 01 TA - bioRxiv PG - 100461 4099 - http://biorxiv.org/content/early/2017/01/15/100461.short 4100 - http://biorxiv.org/content/early/2017/01/15/100461.full AB - Most Genotyping-by-Sequencing (GBS) strategies suffer from high rates of missing data and high error rates, particularly at heterozygous sites. Tunable genotyping-by-sequencing (tGBS®), a novel genome reduction method, consists of the ligation of single-strand oligos to restriction enzyme fragments. DNA barcodes are added during PCR amplification; additional (selective) nucleotides included at the 3’-end of the PCR primers result in more genome reduction as compared to conventional GBS methods. By adjusting the number of selective bases different numbers of genomic sites can be targeted for sequencing. Because this genome reduction strategy concentrates sequencing reads on fewer sites, SNP calls are based on more reads than conventional GBS, resulting in higher SNP calling accuracy (>97-99%) even for heterozygous sites and less missing data per marker. tGBS genotyping is expected to be particularly useful for genomic selection, which requires the ability to genotype populations of individuals that are heterozygous at many loci.