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Gene Expression Profiling of RNA Extracted from FFPE Tissues: NuGEN Technologies’ Whole-Transcriptome Amplification System

  • Leah Turner
  • Joe Don Heath
  • Nurith KurnEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 724)

Abstract

Gene expression profiling of RNA isolated from formalin fixed, paraffin-embedded (FFPE) tissue samples has been historically challenging. Yet FFPE samples are sought-after because of the in-depth retrospective records typically associated with them rendering these samples a valuable resource for translational medicine studies. Extensive degradation, chemical modifications, and cross-linking have made it difficult to isolate RNA of sufficient quality required for large-scale gene expression profiling studies. NuGEN Technologies’ WT-Ovation™ FFPE System linearly amplifies RNA from FFPE samples through a robust and simple whole-transcriptome approach using as little as 50 ng total RNA isolated from FFPE samples. The amplified material may be labeled with validated kits and/or protocols from NuGEN for analysis on any of the major gene expression microarray platforms, including: Affymetrix, Agilent, and Illumina gene expression arrays. Results compare well with those obtained using RNA from fresh-frozen samples. RNA quality from FFPE samples varies significantly and neither sample age nor sample size analysis via gel electrophoresis or the Agilent Bioanalyzer system accurately predict materials suitable for amplification. Therefore, NuGEN has validated a correlative qPCR-based analytical method for the RNA derived from FFPE samples which effectively predicts array results. The NuGEN approach enables fast and successful analysis of samples previously thought to be too degraded for gene expression analysis.

Key words

FFPE Gene expression Isothermal linear amplification 

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Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  1. 1.NuGEN Technologies, Inc.San CarlosUSA

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