FFPE Total RNA Purification Kit (96-well kit)

Product Description

• Extract total RNA (including microRNA) from FFPE samples
• No phenol or chloroform extractions
• Includes DNase for optional on-plate DNA removal
• Isolated RNA is of the highest quality and integrity
• Versatile processing - High throughput isolation of total RNA using either a vacuum manifold or centrifugation
• Isolate a diversity of RNA species - from large mRNA and ribosomal RNA down to microRNA (miRNA) and small interfering RNA (siRNA).
• High yields - Norgen’s FFPE RNA Purification 96-Well Kit allows for the purification of high yields of total RNA.

Norgen’s FFPE RNA Purification Kit provides a rapid method for high throughput isolation and purification of total RNA (including microRNA) from formalin-fixed paraffin-embedded (FFPE) tissue samples. Using formalin to fix tissues leads 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. Norgen’s FFPE RNA Purification Kit provides conditions that allow for the partial reversing of the formalin modifications, resulting in a high quality and yield of RNA. The kit is able to purify all sizes of RNA, from large mRNA and ribosomal RNA down to microRNA (miRNA) and small interfering RNA (siRNA), depending on the age of the FFPE tissue as the degree of fragmentation of the RNA will increase over time. The RNA is preferentially purified from other cellular components without the use of phenol or chloroform.

Purification is based on 96-well column chromatography using Norgen’s proprietary resin as the separation matrix. The process first involves deparaffinization of the FFPE samples through a series of xylene and ethanol washes. Next, the FFPE samples are digested with the provided Proteinase K and Digestion Buffer. Binding Solution and ethanol are then added to the lysate, and the solution is loaded onto a 96-well plate (BIND). Under these conditions only the RNA will bind while most of the contaminants will be removed in the flowthrough. At this point, any remaining traces of genomic DNA can be digested using an optional protocol, allowing for pure RNA samples to be isolated. The bound RNA is then washed (WASH) in order to remove any impurities. Lastly, the purified total RNA is eluted in the provided Elution Buffer or water (ELUTE). Please see the procedure flowchart below.

Workflow

The purified RNA is of the highest integrity, and can be used in a number of downstream applications including qRT-PCR, reverse transcription PCR, primer extension, expression array assays, and microarray analyses.

High Yield and Consistent Recovery of a Diversity of RNA Species. Norgen’s FFPE RNA Purification 96-Well Kit effectively recovers all sizes of RNA including large mRNA to small RNA including microRNAs. Total RNA was isolated from two equal amounts of an FFPE tissue sample using Norgen’s FFPE RNA Purification Kit. The purified RNA was then used as the template in a RT-qPCR for detecting the beta-actin gene (Panel A) and for detecting miR-21(Panel B). As it can be seen, Norgen’s kit successfully and consistently isolated not only large RNA (Panel A) but also microRNA (Panel B), indicating the diversity of RNA species isolated.