Formalin-fixed paraffin-embedded (FFPE) samples introduce unique challenges for gene expression profiling and gene expression quantitation, including chemical modification and fragmentation of RNA molecules. Archival of FFPE samples adds to these challenges through increased RNA degradation over time. As a result, not all FFPE samples contain high-quality RNA.
The Arcturus® Paradise® PLUS Sample Quality Assessment Kit offers a simple solution for evaluating the quality of RNA from formalin-fixed, paraffin-embedded (FFPE) tissues before proceeding with laser microdissection and downstream molecular analysis. The reagents provided in the kit are optimized to enable efficient extraction, isolation, and reverse transcription of total RNA from FFPE tissue scrapes. The RNA quantity and quality is then measured using quantitative real-time PCR (Figure 1).
The RNA quantity and quality are assessed by the presence of intact RNA for a specific message. Two sets of β-actin gene-specific primers designed to amplify short segments of DNA within the β−actin target sequence are used to estimate the amount of RNA in a given sample in relation to β-actin content using the real-time PCR technique. The RNA quantity derived from the 3′ primer set is used to quantitate the RNA in the FFPE tissue scrape. The ratio of the RNA yield obtained from both sets of PCR primers is the 3'/5' ratio and is used as an indication of RNA quality.
|Figure 1. Real-time results identify quality samples to use in further experimentation.
The Paradise® PLUS Reagent System is the only available solution that provides all the components necessary to prepare RNA from FFPE tissue samples for gene expression analysis. To help ensure successful microarray results, the system includes reagents optimized for exceptional recovery of RNA and superior RNA amplification.
The Paradise® PLUS Reagent System is validated as part of the Arcturus® Complete Systems for Microgenomics®, an integrated solution for utilizing small quantities of RNA for gene expression analysis, and features Arcturus Laser Capture Microdissection (LCM) instruments to procure pure cell populations. Arcturus' Systems for Microgenomics® enable accurate and sensitive microarray assays that reveal molecular signatures otherwise obscured in whole tissue samples or cell mixtures.
For Research Use Only. Not for use in diagnostic procedures.