Rapid, Low-plex Genotyping of Wheat and Other Cereals

Eureka™ Genotyping Solution is a novel technology that has utility in plant breeding and regulatory aspects. It allows for two-day turnaround time, from sample to genotype calls for catalog or fully customizable panels of a few 100s up to 3000 variants (substitutions or INDELS). The Eureka™ genotyping assay is a ligation-dependent polymerase chain reaction made ready for high through put sequencing. It consists of probe triplets in solution, where for each variant there are two left side probes (one for each allele) immediately followed by a third right side probe. The probes are hybridized to target on the input DNA and a ligation step locks the correctly hybridized left and right probes. Interrogation site bar codes are contained within the ligation probes and sample index bar codes are added during the post-ligation amplification step. Amplicons from all the samples are pooled and short-cycle sequence data is generated from the prepared libraries. The Eureka™ Analysis Suite software is used to tabulate the number of reads that contain each combination of sample, locus, and allele bar code (as appropriate). The genotype of each sample for each locus is inferred from statistical analysis of the tabulated reads. The Eureka Genotyping Solution is affordable, flexible, accurate and robust. Results for Eureka Genotyping Panels developed for Hordeum vulgare, Glycine max, Zea mays and the genetically more complex Triticum aestivum will be discussed