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The Arcturus® Paradise® PLUS Reagent System enables unprecedented gene expression analysis of millions of previously inaccessible samples. The Paradise® PLUS Reagent System is an integrated solution for gene expression studies from formalin-fixed, paraffin-embedded (FFPE) tissues, including reagents for tissue staining, RNA isolation, and RNA amplification. Amplify RNA from as little as 5 ng of fixed RNA using methods that have been specially optimized to overcome the challenges of cross-linked templates.
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Two versions of the kit include:
Currently, tens of millions of tissue samples with known outcomes are available in tissue archives. The Paradise® PLUS Reagent System enables microarray and qRT-PCR analysis of these previously inaccessible samples, allowing retrospective studies to accelerate discoveries.
Only six steps are required to take you from sample to results (Figure 1).
Figure 1. Six steps to achieve gene expression results. |
The Paradise® PLUS Reagent System is compatible with all major microarray platforms. Robust and reproducible results are achieved using tools and techniques commonly employed in today’s research laboratories (Figures 2–5).
Figure 2. Paradise® PLUS reproducibility on GeneChip® Human Genome X3P array (Affymetrix). Amplification was performed on FFPE breast cancer samples. The double-stranded cDNA generated was used as template for labeling during in vitro transcription (IVT). Duplicate samples were hybridized to GeneChip® microarrays, resulting in intensity correlation of R=0.983. |
Figure 3. Paradise® PLUS reproducibility on cDNA arrays. Two-round amplifications were performed on independent laser capture microdissected FFPE tissue cells. Amplified aRNA was labeled during reverse transcription and hybridized to two cDNA arrays, resulting in correlation of gene expression ratios, R= 0.993 (log2 ratios). |
Figure 4. Paradise® PLUS microarray reproducibility. Two-round amplifications were performed on independent laser capture microdissected human breast cells. Labeled aRNA was hybridized to custom-designed Agilent microarrays, resulting in correlation of gene expression ratios, R=0.986 (log2 ratios). Additionally, amplification across a variety of tissues is robust, providing microgram quantities of aRNA suitable for labeling and hybridization to microarrays. |
Figure 5. Samples processed using the Paradise® PLUS Reagent System. Starting with 5,000–10,000 laser capture microdissected cells produces amplified RNA sufficient for duplicate microarray hybridizations. |
Not all FFPE samples contain high-quality RNA. Identify the best samples for your research studies using a simple real-time PCR technique, and then proceed with confidence using these prequalified samples (See Figure 6.)
Figure 6. 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 formalin-fixed, paraffin-embedded 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. This system 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.