Efficient Creation of Long-Template Libraries for Next-Generation Sequencing

DESCRIPTION provided by applicant The advent of next generation sequencing NGS technologies has allowed researchers to make unprecedented progress in the analysis of genomes the results of which have profound implications in the realms of human health agricultural productivity and natural history The ability to fully characterize genomes remains unrealized however due in large part to our inability to reconcile sequence information over long distances Short fragmented DNA libraries and their subsequent alignment to reference genomes while highly useful in identifying single nucleotide variants SNV make de novo genome assembly and genomic structural analysis difficult Long template NGS sequencing offers a potential solution but suffers from complex and expensive library preparation strategies Such strategies frequently require significant amounts of input DNA due to complicated and inefficient manipulation steps We have proposed an approach for creating long template libraries that addresses these issues with a simple efficient and relatively inexpensive method involving the innovative pairing of long dumbbell templates with solid phase rolling circle replication The successful development of this technology would greatly enhance the research communityandapos s ability to interrogate genomes particularly those with large structural variation which likely plays a significant role in oncogenesis and tumor progression PUBLIC HEALTH RELEVANCE The study and sequencing of human DNA has lead to amazing discoveries in forensics history and medicine As of the methods associated with preparing a DNA sample for sequencing are prone to inefficiency waste and error all of which increase the cost of the process The research proposed here is oriented toward significantly improving the methods of sample preparation which will lead to improved efficiency accuracy and reduced costs to sequence DNA thereby making the technology more accessible to more people