Project Summary/Abstract Expanding our understanding of the genetic contributions and etiologies of congenital disabilities and childhood cancer impacts those affected directly and contributes significantly to related research in adult conditions. This proposal builds on the established HudsonAlpha-St. Jude Genome Sequencing Center (HASJ-GSC) and expands its capabilities to interrogate and integrate data produced by an efficient and experienced genomics resource. The program aims to generate and analyze high-quality sequence and variant data from the most pediatric cancer and congenital disability cohorts possible with available funds over three years. We present here the innovation, capabilities, and experience of the HASJ-GSC to produce data for the Gabriella Miller Kids First Research Program that will be accessible and available to the research community and leveraged to its maximal impact for years to come. We also present a technology expansion and cost reduction plan that increases the number of cases sequenced and analyzed each year. The HASJ-GSC will generate exceptional quality whole-genome sequence and variant data for all samples. The pediatric cancer samples will also be analyzed with RNASeq and whole-exome sequencing. The combined genome, exome, and RNASeq will provide as much resolution as possible toward understanding the genetic and functional genomic changes observed in pediatric cancer. The HASJ-GSC will also support a comprehensive collection of additional methodologies such as long-read DNA and RNA sequencing, functional genomic analyses, and advanced data analysis. These technologies will expand the resolution and types of investigations provided to X01 investigators. The HASJ- GSC will also provide data storage with fast, reliable, and efficient data access, sharing, and reporting for the X01 project researchers. Finally, the HASJ-GSC will support efficient sequence and variant data submission to the Gabriella Miller Kids First Data Resource Center and facilitate submission to the appropriate public databases.