Amplicon-Based Targeted Next-Generation Sequencing of Formalin-Fixed, Paraffin-Embedded Tissue

Eric Strengman, Francoise A.S. Barendrecht-Smouter, Carmen de Voijs, Paula de Vree, Isaac J. Nijman, Wendy W.J. de Leng

Research output: Chapter in Book/Report/Conference proceedingChapterAcademicpeer-review

Abstract

Next-generation sequencing (NGS) is rapidly becoming the method of choice for mutation analysis in both research and diagnostics. The benefit of targeted NGS compared to whole-genome and whole-exome sequencing is that smaller amounts of input material can be used as well as qualitatively suboptimal tissue samples, like formalin-fixed, paraffin-embedded archival tissue.Here, we describe the protocol for targeted next-generation sequencing using the Ion Torrent PGM platform in combination with Ion Ampliseq NGS gene panels for formalin-fixed, paraffin-embedded tissues. Both the manual and the automated workflow are described as well as the bioinformatics for data analysis.

Original languageEnglish
Title of host publicationTumor profiling
Subtitle of host publicationmethods and protocols
EditorsSarah S. Murray
Place of PublicationNew York
PublisherSpringer Science + Business Media
Chapter1
Pages1-17
Number of pages17
Volume1908
ISBN (Electronic)978-1-4939-9004-7
ISBN (Print)978-1-4939-9002-3
DOIs
Publication statusPublished - 1 Jan 2019

Publication series

NameMethods in Molecular Biology
Volume1908
ISSN (Print)1064-3745

Keywords

  • Ampliseq
  • Formalin-fixed paraffin-embedded
  • Mutation analysis
  • Next-generation sequencing
  • Targeted sequencing
  • Variant calling

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