Next‐Generation Sequencing Library Preparation from FFPE Tissue Samples. Issue 1 (4th January 2016)

• Record Type: Journal Article
• Authors: Wolf, Kathrin; O'Neil, Dominic; Schroeer, Stefanie; Fang, Nan
• Editors: Ausubel, Frederick M.; Brent, Roger; Kingston, Robert E.; Moore, David D.; Seidman, J.G.; Smith, John A.; Struhl, Kevin
• Is Part Of: Current protocols in molecular biology. Volume 113:Issue 1(2016)
• Page Start: 7.24.1
• Page End: 7.24.14
• Publication Date: 2016-01-04
• Subjects: formalin‐fixed paraffin‐embedded tissue -- next‐generation sequencing -- fragmentation -- library preparation -- illumina; Molecular biology -- Technique -- Periodicals; Molecular biology -- Laboratory manuals; Molecular Biology -- methods; Biologie moléculaire -- Technique; Biologie moléculaire -- Manuels de laboratoire; Molecular biology; Molecular biology -- Technique; Laboratory Manual; Electronic reference sources; Laboratory manuals; 572.8028
• DOI: 10.1002/0471142727.mb0724s113
• Languages: English
• ISSNs: 1934-3639

Abstract

Abstract: Formalin‐fixed, paraffin‐embedded (FFPE) tissue samples represent an invaluable biobank for retrospective cancer research with molecular methods such as real‐time PCR and next‐generation sequencing (NGS). However, the usage of FFPE material in NGS approaches involves several challenges associated with the limited quantity and quality of DNA. This protocol describes how to purify DNA from FFPE material and how to prepare it for downstream NGS workflows including fragmentation, size selection, and library preparation for Illumina MiSeq/HiSeq sequencing. © 2016 by John Wiley & Sons, Inc.