Norgen’s Total RNA Purification Plus Micro Kit provides a rapid and sensitive method for the isolation and purification of total RNA from small input amounts of cultured animal cells, tissue samples, and microdissected samples including laser-capture microdissection (LCM). The kit purifies all sizes of RNA, from large mRNA and ribosomal RNA down to microRNA (miRNA) and small interfering RNA (siRNA). Genomic DNA is removed from the sample using a Genomic DNA Removal column, and the RNA is preferentially purified from other cellular components such as proteins, without the use of phenol or chloroform. The purified RNA is of the highest integrity, and can be used in a number of downstream applications including real time PCR, reverse transcription PCR, Northern blotting, RNase protection and primer extension, and expression array assays.

Norgen’s Purification Technology

Purification is based on spin column chromatography using Norgen’s proprietary resin as the separation matrix. The RNA is preferentially purified from other cellular components such as proteins without the use of phenol or chloroform. The process involves first lysing the cells or tissue of interest with the provided Buffer RL (please see the flow chart on page 4). The DNA is then captured on a Genomic DNA Removal Column. Ethanol is then added to the flowthrough of the DNA removal step, and the solution is loaded onto an RNA Purification Micro Column. Norgen’s resin binds RNA in a manner that depends on ionic concentrations. Thus only the RNA will bind to the column, while the contaminating proteins will be removed in the flowthrough or retained on the top of the resin. The bound RNA is then washed with the provided Wash Solution A in order to remove any remaining impurities, and the purified total RNA is eluted with the Elution Solution A. The special design of the micro spin-column allows a small elution volume of as little as 20 μL. The purified RNA is of the highest integrity, and can be used in a number of downstream applications.