title: Generating Exome Enriched Sequencing Libraries from Formalin-Fixed, Paraffin-Embedded Tissue DNA for Next-Generation Sequencing.
abstract: This unit describes a technique for generating exome-enriched sequencing libraries using DNA extracted from formalin-fixed paraffin-embedded (FFPE) samples. Utilizing commercially available kits, we present a low-input FFPE workflow starting with 50 ng of DNA. This procedure includes a repair step to address damage caused by FFPE preservation that improves sequence quality. Subsequently, libraries undergo an in-solution-targeted selection for exons, followed by sequencing using the Illumina next-generation short-read sequencing platform. © 2017 by John Wiley & Sons, Inc.
authors.affiliation: Center for Inherited Disease Research, McKusick-Nathans Institute of Genetic Medicine, Johns Hopkins University, Baltimore, Maryland.
authors.forename: Beth A
authors.initials: BA
authors.lastname: Marosy
authors.affiliation: Center for Inherited Disease Research, McKusick-Nathans Institute of Genetic Medicine, Johns Hopkins University, Baltimore, Maryland.
authors.forename: Brian D
authors.initials: BD
authors.lastname: Craig
authors.affiliation: Center for Inherited Disease Research, McKusick-Nathans Institute of Genetic Medicine, Johns Hopkins University, Baltimore, Maryland.
authors.forename: Kurt N
authors.initials: KN
authors.lastname: Hetrick
authors.affiliation: Center for Inherited Disease Research, McKusick-Nathans Institute of Genetic Medicine, Johns Hopkins University, Baltimore, Maryland.
authors.forename: P Dane
authors.initials: PD
authors.lastname: Witmer
authors.affiliation: Center for Inherited Disease Research, McKusick-Nathans Institute of Genetic Medicine, Johns Hopkins University, Baltimore, Maryland.
authors.forename: Hua
authors.initials: H
authors.lastname: Ling
authors.affiliation: Center for Inherited Disease Research, McKusick-Nathans Institute of Genetic Medicine, Johns Hopkins University, Baltimore, Maryland.
authors.forename: Sean M
authors.initials: SM
authors.lastname: Griffith
authors.affiliation: Center for Inherited Disease Research, McKusick-Nathans Institute of Genetic Medicine, Johns Hopkins University, Baltimore, Maryland.
authors.forename: Benjamin
authors.initials: B
authors.lastname: Myers
authors.affiliation: National Human Genome Research Institute, National Institutes of Health, Bethesda, Maryland.
authors.forename: Elaine A
authors.initials: EA
authors.lastname: Ostrander
authors.affiliation: Program in Prostate Cancer Research, Division of Public Health Sciences, Fred Hutchinson Cancer Research Center, Seattle, Washington.
authors.forename: Janet L
authors.initials: JL
authors.lastname: Stanford
authors.affiliation: Medical Genomics and Metabolic Genetics Branch, National Human Genome Research Institute, Bethesda, Maryland.
authors.forename: Lawrence C
authors.initials: LC
authors.lastname: Brody
authors.affiliation: Center for Inherited Disease Research, McKusick-Nathans Institute of Genetic Medicine, Johns Hopkins University, Baltimore, Maryland.
authors.forename: Kimberly F
authors.initials: KF
authors.lastname: Doheny
mesh_terms: D004247:DNA / Q000235:genetics*; D059472:Exome / Q000235:genetics*; D005557:Formaldehyde; D015723:Gene Library; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D006801:Humans; D010232:Paraffin; D016612:Paraffin Embedding; D016707:Tissue Fixation; D000073359:Exome Sequencing
chemical_list: D005557:Formaldehyde; D010232:Paraffin; D004247:DNA
keywords: DNA library preparation; DNA repair; FFPE; next-generation sequencing; whole exome sequencing