WT-Ovation™ FFPE System V2
The NuGEN WT-Ovation™ FFPE System V2 is a whole transcriptome amplification system enabling researchers to conduct global gene expression analysis on the vast archives of small and degraded RNA derived from FFPE samples. The system is comprised of reagents and a protocol required for amplification of as little as 50 ng of total FFPE RNA. The protocol takes just six hours, and can be used for qPCR, sample archiving, fragmentation, and labeling.
The amplified cDNA can be fragmented and labeled in less than two hours for GeneChip® 3’ expression array analysis using NuGEN’s FL-Ovation™ cDNA Biotin Module V2.
For analysis using Affymetrix GeneChip® Exon and Gene ST arrays, the amplified cDNA can be used with the WT-Ovation Exon Module, then fragmented and labeled using the FL-Ovation™ cDNA Biotin Module V2.
For analysis on Agilent arrays, the amplified cDNA can be fragmented and labeled using NuGEN’s FL-Ovation™ cDNA Fluorescent Module.
The WT-Ovation™ FFPE System V2 provides optimized reagent mixes and a protocol to process 12 or 60 RNA samples (Cat.# 3400-12, 3400-60).
Principle Components Analysis (PCA) of colon tumor and normal adjacent tissue (NAT): Targets were prepared from RNA extracted from formalin fixed paraffin embedded tissue (FFPE) from one donor (pink) and fresh frozen tissue from a second donor (blue). Each sample was amplified in quadruplicate and hybridized to Affymetrix HGU133A_2.0 GeneChip® arrays. PCA was performed using Partek Genomics Suite software. The green and blue ellipses (NAT and Tumor, respectively) define the boundary of two standard deviations from the centroid of each cluster indicating a statistically significant separation of samples based on the disease state of the tissue. This demonstrates that the amplification system maintains the integrity of the biological data.
