Targeted Next –Generation Sequencing for Canine Genotyping Using Fluidigm® D3™ Assay design and the Access Array™ System

The advantages of rapidly increasing throughput and decreasing cost in next-generation technology make it more practicable in genetic variation discovery using the next generation sequencing (NGS) data. The targeted genotyping approaches are ideal for Single Nucleotide Polymorphism (SNP) in plant and animal functional genomics studies after the markers have been discovered.  Targeted assay design and sample preparation are the two main challenges to implement this approach. As a proof of concept experiment, we used Fluidigm D3 Assay designed a set of Access Array target-specific primers. The sequence ready library is prepared in conjunction with Fluidigm Access Array Barcode Library and Access Array system. The genotyping results for all samples were obtained in a single sequencing run. This experiment demonstrates a simplified NGS library preparation method that can be utilized in the various genetic studies to achieve higher sample throughput and more accurate data.