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 75 µL of the provided Elution Buffer or water (ELUTE). Please see the procedure flowchart to the right.
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
FEATURES AND BENEFITS
- High quality and integrity of the isolated RNA - The purified total RNA is of the highest quality and integrity, and can be used in any sensitive downstream applications.
- Isolate a diversity of RNA species - All RNA species can be isolated, 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.
- No phenol:chloroform extractions - Total RNA is isolated from FFPE tissue samples without the use of harmful chemicals such as phenol or chloroform.
- Versatile processing - High throughput isolation of total RNA using either a vacuum manifold or centrifugation.