Generating exome enriched sequencing libraries from formalin-fixed, paraffin-embedded tissue DNA for next-generation sequencing

Beth A. Marosy, Brian D. Craig, Kurt N. Hetrick, P. Dane Witmer, Hua Ling, Sean M. Griffith, Benjamin Myers, Elaine A. Ostrander, Janet L. Stanford, Lawrence C. Brody, Kimberly F. Doheny

Research output: Contribution to journalArticlepeer-review

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.

Original languageEnglish (US)
Pages (from-to)18.10.1-18.10.25
JournalCurrent protocols in human genetics
Volume2017
DOIs
StatePublished - Jan 1 2017

Keywords

  • DNA library preparation
  • DNA repair
  • FFPE
  • Next-generation sequencing
  • Whole exome sequencing

ASJC Scopus subject areas

  • Genetics
  • Genetics(clinical)

Fingerprint

Dive into the research topics of 'Generating exome enriched sequencing libraries from formalin-fixed, paraffin-embedded tissue DNA for next-generation sequencing'. Together they form a unique fingerprint.

Cite this