Targeted long-read sequencing sample preparation using Cas9 nucleases

Project Summary DNA sequencing is increasingly being used in clinical research in genetic disease and oncologyCurrently most clinical sequencing is carried out using short read targeted sequencing methods to detect mutations in protein coding regions of the genomeHowevershort read sequencing methods are not well suited to detection of alterations involving DNA segments greater thanbase pairs in lengthnor can they detect the arrangement of sequence polymorphisms that are more than a few hundred bases apart on a chromosomeSuch long range genomic analyses are becoming increasingly importantand new long read sequencing technologies have been developed that can address these technical problemsHoweverthe new long read sequencing methods are roughlyfold more expensive than commonly used short read sequencing methodsOur proposal seeks to develop an instrument system that can isolate specific long genomic DNA fragmentstomillion base pairs in lengthfrom biological samplesand thereby provide a new economical approach for targeted long read sequencing sample preparationThe proposed system is intended for robusthigh sample throughputwalk away automated processing in high volume genome centers and clinical laboratories Project Narrative Current DNA sequencing used in clinical research is focused on detecting single base mutations in proteincoding regions of the genomeHowevermuch genetic variation in inherited genetic disease and cancer involves alterations of large DNA regionsand such alterations are not detected by current short read clinical sequencing methodsThis application proposes development of a new sample preparation instrument that will enable new long read sequencing methods to be used cost effectively in clinical research