The Applied Biosystems® Arcturus® Paradise® PLUS Whole Transcript RT (WT-RT) Reagent System from Life Technologies provides an efficient and reliable solution for purifying RNA from archived formalin-fixed, paraffin-embedded (FFPE) tissue samples followed by reverse transcription of the whole transcript for use in real-time PCR.
Key Product Features
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.
The Paradise® PLUS WT-RT Reagent System was specifically developed and specially optimized to provide high-quality RNA and robust reverse transcription (RT) from highly fragmented RNA obtained from archived FFPE samples over 20 years old (Figure 2).
- Simple workflow—minimal sample handling reduces chance of contamination (Figure 3)
- Efficient RNA extraction and isolation—helps save time and minimizes sample loss
- Optimized RT reactions—generate cDNA representing the entire length of the transcript
- Accurate transcript analysis—results represent high, medium, and low-abundance expressing genes
Optimized RNA Extraction and Isolation from Archived FFPE Tissues
Table 1 shows a comparison of total RNA yields using the Paradise® PLUS WT-RT Reagent System and a similar commercially available protocol. The average yield of the three replicate isolations illustrates the abundance of total RNA made available using the Paradise® PLUS WT-RT Reagent System. The Paradise® PLUS WT-RT Reagent System produces abundant total RNA for reverse transcription and subsequent qPCR analysis, up to three times more total RNA than obtained when a similar commercially available product was used for the same sample.
Table 1. Comparison of total RNA yields using the Paradise® PLUS WT-RT Reagent System and a similar commercially available protocol.
| Sample ID | Sample Age | 260/280 | RNA Isolation Protocol | Avg. Yield (ng) |
|---|---|---|---|---|
| 9055 | 1 year | WT-RT | 1.94 | 7592.8 |
| Vendor A | 2.05 | 2086.3 | ||
| 8090 | 10 years | WT-RT | 1.95 | 784.4 |
| Vendor A | 2.45 | 247.4 |
Superior Amplification Enabled Over a Wide Range of Targets
The sensitivity of the Paradise® PLUS WT-RT Reagent System was compared to similar reagent products available in the market that generate cDNA for use in real-time PCR reactions. Improved Ct values were seen for all high, medium, and low expressing genes when the Paradise® PLUS product was used (Figure 2). Exon-spanning primers at varying distances from the 3’-end of the transcript (500 bp to 5 kb) were used to show increased sensitivity and improved transcription success compared to other commercially available products. The Paradise® PLUS WT-RT Reagent System successfully amplified all genes, because the reagents provided in the kit are optimized to work together, helping you save time and effort while producing high-quality results across the entire transcript.
| Figure 2. Improved Ct values were seen for all high, medium, and low expressing genes when the Paradise® PLUS product was used. |
Complete Systems for Microgenomics®
The Paradise® PLUS Reagent Systems 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.