Figure 1. High Quality and Yield of Total RNA. Norgen's FFPE RNA Purification Kit Dx isolates FFPE RNA that exceeds both yield and quality of competitors. Total RNA was isolated from one slice of hamster FFPE kidney section (20 micron thickness) using Norgen's FFPE RNA Purification Kit Dx and a leading competitor’s kit. One microliter of the 50 µL purified RNA was then resolved on an Agilent 2100 BioAnalzyer using an RNA Nano 6000 chip. As it can be seen, Norgen not only isolated higher yields of total RNA, but the RNA was also of a higher quality as evidenced by the higher RIN values obtained with Norgen's RNA.
Figure 2. Higher Yield of FFPE RNA Isolated by Norgen's FFPE RNA Purification Kit Dx. Norgen's FFPE RNA Purification Kit Dx isolates FFPE RNA that exceeds the yield of competitors. Total RNA was isolated from one slice of hamster FFPE kidney section (20 micron thickness) using Norgen's FFPE RNA Purification Kit Dx and two leading competitor's kits. Eighteen isolations were performed for each product. The graph demonstrates the mean yield of RNA according to spectrophotometry for 18 sample replicates. Vertical bars represent the standard deviation. Norgen's kit consistently purified total RNA with a higher yield than for those obtained using the market competitor's kits.
Figure 3. High Yield of a Diversity of RNA Species. Norgen's FFPE RNA Purification Kit Dx effectively recovers all sizes of RNA from large mRNA to small RNA including microRNAs. Total RNA was isolated from one slice of hamster FFPE kidney section (20 micron thickness) using Norgen's FFPE RNA Purification Kit Dx and a leading competitor’s kit. Five microliters of the 50 µL purified RNA was then used as the template in a 20 µL reverse transcription reaction with oligo dT primer or miR-21 stem-loop reverse primer. Two microliters of the RT reaction was then used in a 15 µL qPCR reaction for detecting the beta-actin gene (Panel A) and for detecting miR-21(Panel B), respectively. In both graphs the blue lines correspond to Norgen isolated-RNA and the green lines correspond to competitor-isolated RNA. As it can be seen, Norgen's kit isolated higher yields of RNA in both cases, as indicated by the lower Ct values of the blue lines. Also, Norgen's kit successfully isolated not only large RNA (Panel A) but also microRNA (Panel B), indicating the diversity of RNA species isolated.
Figure 4. Excellent Performance in Downstream Application such as RT-qPCR. Norgen's FFPE RNA Purification Kit Dx effectively recovers all sizes of RNA, from large mRNA to small RNA including microRNA, that perform effectively in sensitive applications such as RT-qPCR. Total RNA was isolated from equal amounts of an FFPE tissue sample using Norgen's FFPE RNA Purification Kit and two market competitors. The graph demonstrates the mean delta Ct value for 18 sample replicates. Vertical bars represent the standard deviation. Norgen's kit consistently isolates RNA of a higher yield and quality than that obtained by the two leading market competitors as indicated by the larger delta Ct values. Not only were the large RNA species detected including rRNA and mRNA, but small RNA species (microRNA) were also detected from the sample, indicating the diversity of RNA species isolated.
5 slices of < 20 µm thick paraffin slices
25 mg of unsectioned block
Average Yield
Variable due to age of paraffin blocks
~2-3 µg of Total RNA per 1 mg of fresh FFPE hamster kidney
* Time required for purification varies by length of Proteinase K incubation and formalin crosslink-reversal
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Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. The DNAse I and Proteinase K should be stored at -20°C upon arrival. These reagents should remain stable for at least 2 years in their unopened containers.
