Cytoplasmic & Nuclear RNA Purification Kit # 21000, 37400
Norgen’s Cytoplasmic & Nuclear RNA Purification Kit provides a rapid method for the isolation
and purification of both cytoplasmic and nuclear RNA from cultured animal cells and small tissue
samples. In certain circumstances it is desirable to be able to isolate fractionated RNA as
opposed to total RNA. For example, it may be preferable to isolate only mature, cytoplasmic
RNA for some studies on expression profiling. Alternatively it may be desirable to isolate nuclear
RNA in order to investigate and study pre-processed (non-spliced) RNA. Furthermore, this kit
can be used to isolate RNA for downstream applications where it is necessary to avoid DNA
contamination, since the cytoplasmic fraction has been shown to be free of all traces of genomic
DNA. Norgen’s Cytoplasmic & Nuclear RNA Purification Kit can be used to isolate all sizes of
RNA from the cytoplasmic and nuclear RNA fractions, including all small RNA species. The kit is
supplied with sufficient reagents to perform either 50 cytoplasmic RNA preparations or 25
cytoplasmic and 25 nuclear RNA preparations.
Norgen’s Purification Technology
Purification is based on spin column chromatography using Norgen’s proprietary resin as the
separation matrix. The cytoplasmic RNA is preferentially purified from the nuclear RNA and 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 Lysis Buffer J (please see the
flow chart on page 4). The lysate is then separated through centrifugation, with the supernatant
containing the cytoplasmic RNA and the pellet containing the nuclear RNA. Buffer SK and
ethanol are then added to the desired fraction, and the solution is loaded onto a spin-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 RNA is eluted with the Elution
Buffer E. 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.
