Prelude™ FFPE RNA Isolation Module
The Prelude FFPE RNA Isolation Module provides a fast and easy method for isolating total RNA from formalin-fixed paraffin-embedded (FFPE) tissue samples. The protocol has been optimized to recover total RNA for input to NuGEN's FFPE sample preparation solutions including Ovation® FFPE WTA System (Part No. 3403) and Ovation RNA-Seq FFPE System (Part No. 7150).
Use of formalin to fix tissues can lead to crosslinking of the RNA and proteins, and the process of embedding the tissue samples can also lead to fragmentation of the RNA over time. NuGEN's Prelude FFPE RNA Isolation Module provides conditions that allow for the partial reversing of the formalin-induced modifications, resulting in a high quality and yield of RNA for downstream processing. The total RNA is preferentially purified from other cellular components without the use of phenol or chloroform. The purified total RNA can be used in a number of downstream applications including qPCR, microarray analyses, and sequencing (RNA-Seq).
The Prelude FFPE RNA Isolation Module (Part No. 1410-50) contains enough reagents and supplies to perform fifty total RNA isolations from FFPE samples.
| FFPE Sample | Total RNA Input | cDNA Yield | %P | SF | Actin | GAPDH |
|---|---|---|---|---|---|---|
| Human Liver (2 year old) |
50 ng | 8.8 ug | 55.9 | 4.75 | 5.24 | 3.66 |
| Human Lung (8 year old) |
50 ng | 6.1 ug | 52.9 | 3.43 | 5.1 | 3.3 |
Array Performance Metrics with Total RNA isolated using the Prelude FFPE RNA Isolation Module: Total RNA was isolated from either Human liver or lung FFPE samples that were stored for 2 years or 8 years, respectively. cDNA targets were prepared from triplicate (liver) or duplicate (lung) isolations using the Ovation FFPE WTA System (Part No. 3403). The amplified cDNA was fragmented and labeled using the Encore Biotin Module (Part No. 4200), and hybridized to GeneChip® Human Genome U133A 2.0 arrays. The data shows good %P values with low Actin and GAPDH 3' to 5' ratios, which illustrates high quality data both in terms of transcript coverage and representation of low abundant transcripts.
Ordering Information
| Product Name | Part No. | No. of Reactions |
|---|---|---|
| Prelude FFPE RNA Isolation Module | 1410-50 | 50 |
| Related Products | ||
| Ovation FFPE WTA System | 3403-12 | 12 |
| 3403-60 | 60 | |
| 3403-A01 | 96 | |
| Encore Biotin Module | 4200-12 | 12 |
| 4200-60 | 60 | |
| 4200-A01 | 96 | |
| Ovation RNA-Seq FFPE System | 7150-08 | 8 |
