Generating Exome Enriched Sequencing Libraries from Formalin-Fixed, Paraffin-Embedded Tissue DNA for Next-Generation Sequencing

Curr Protoc Hum Genet. 2017 Jan 11:92:18.10.1-18.10.25. doi: 10.1002/cphg.27.

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.

Keywords: DNA library preparation; DNA repair; FFPE; next-generation sequencing; whole exome sequencing.

MeSH terms

  • DNA / genetics*
  • Exome / genetics*
  • Exome Sequencing*
  • Formaldehyde*
  • Gene Library*
  • High-Throughput Nucleotide Sequencing / methods*
  • Humans
  • Paraffin
  • Paraffin Embedding*
  • Tissue Fixation*

Substances

  • Formaldehyde
  • Paraffin
  • DNA