Exome Sequencing: Applications From the Lab Bench to the Clinic

摘要: Since the initial report of targeted-enrichment (Albert et al, 2007) we have been evolving design and utility capture reagents methods, while taking advantage parallel advances in sequencing platforms. New exome designs target a comprehensive set coding exons from 6 different gene databases, as well computationally predicted non-coding elements: regulatory regions, conserved UTRs. Library automation, reduction DNA input samples, hybridization multiplexing application faster read mapping tools such BWA, together allow rate >4,300 libraries/captures per month, with >40,000 regional libraries completed to date. In addition, fully integrated informatics analysis pipeline (Mercury), supports all aspects data flow production on instrument annotated variant calls (SNPs small Indels). These laboratory methods pipelines hardened at Human Genome Sequencing Center (HGSC) now applied toward clinical sequencing. Through joint collaboration between Medical Genetics Laboratories (MGL) Department Molecular Genetics, interpretation are provided through CAP/CLIA certified Whole Laboratory (WGL). To date, WGL has 650 patient samples final for over 450 patients causative deleterious mutations identified 25% cases. Performance maintained high standard 95% bases represented 20X coverage. Overall performance metrics, LIMS support, validation environment will be presented.