Novel method to capture long, intact DNA for NGS applications

Project Summary Next generation sequencing platforms have fundamentally altered genetic diagnostics and genomic research by providing massive amounts of sequence data in a low costhigh throughput formatA major drawback of short sequence read platforms is their inability to resolve complex genomic regionsLong sequence read platforms can overcome this issuebut existing methods for sequence enrichment are unable to provide the large fragments required to analyze confounding sequence elements at important locisuch as determination of structural and copy number variation in complex genomic regions associated with human diseaseThe goal of this Phase I proposal is to demonstrate the feasibility of using Varigenandapos s amplification freeDNATrapmethod to isolate complex and problematic genomic regions ofkb from humanrat and macaque genomesThis platform independentsequence specific enrichment technology will provide full length epigenetic sequence analysis of these regionsproviding the basis to replace multiple molecular assays presently used for genetic testingDevelopment of the proposed technology will enable isolation and full length sequencing of any genomic regionwhether simple or complexproviding valuable long range haplotype resolved information of disease genes for research or diagnostic applicationsLong term goals of this project in Phase II are to develop and validate diagnostic tests for the disease regions studied in Phase I Narrative Current methods for diagnostic DNA sequencing are unable to resolve complex genomic locibecause they cannot target the largecomplex fragments required to span confounding sequencesnecessitating the use of multiple assays to accurately diagnose disease genesWe will fill this unmet need with the development of an innovative tool to rapidly and affordably capture and sequence full length targeted segments up tokb of human and microbial genomes associated with diseaseThis achievement will significantly improve the ability of researchers and clinicians to obtain complete and accurate characterization and diagnosis of complex genomic variations associated with human disease