Open AccessGenerating Exome Enriched Sequencing Libraries from Formalin‐Fixed, Paraffin‐Embedded Tissue DNA for Next‐Generation Sequencing
Author(s) -
Marosy Beth A.,
Craig Brian D.,
Hetrick Kurt N.,
Witmer P. Dane,
Ling Hua,
Griffith Sean M.,
Myers Benjamin,
Ostrander Elaine A.,
Stanford Janet L.,
Brody Lawrence C.,
Doheny Kimberly F.
Publication year - 2017
Publication title -
current protocols in human genetics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.282
H-Index - 30
eISSN - 1934-8258
pISSN - 1934-8266
DOI - 10.1002/cphg.27
Subject(s) - dna sequencing , exome sequencing , exome , computational biology , illumina dye sequencing , dna , biology , workflow , microbiology and biotechnology , genetics , computer science , gene , mutation , database
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.