Cytoplasmic & Nuclear RNA Purification Kit
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 as little as 45 minutes. In certain circumstances it is desirable 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, or nuclear RNA in order to study pre-processed 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.
Purification is based on spin column chromatography using Norgen’s proprietary resin as the separation matrix. Briefly, the process involves first lysing the cells or tissue of interest with the provided Lysis Solution. The lysate is then separated through centrifugation, with the supernatant containing the cytoplasmic RNA and the pellet containing the nuclear RNA. Binding solution and ethanol are then added to the desired fraction, and the solution is loaded onto a spin-column (BIND 1 and BIND 2). Under these conditions 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 in order to remove any remaining impurities (WASH 1 and WASH 2). Lastly, the desired purified RNA is eluted into 50 µL of the provided Elution Buffer or water (ELUTE 1 and ELUTE 2). 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 real time PCR, reverse transcription PCR, Northern blotting, RNase protection and primer extension, expression array assays, miRNA cloning and Next Generation Sequencing.
FEATURES AND BENEFITS:
- Fast and easy processing – Rapid spin-column format allows for the processing of 10 samples in 45 minutes.
- No genomic DNA contamination in cytoplasmic fraction – Purified cytoplasmic RNA can be used directly in RT-PCR reactions with no DNase treatment required.
- No phenol:chloroform extractions- Cytoplasmic and nuclear RNA is isolated without the use of harmful chemicals such as phenol or chloroform.
- Isolate a diversity of RNA species – All nuclear and cytoplasmic RNA species can be isolated, from large mRNA species down to microRNA (miRNA) and small interfering RNA (siRNA).
- High quality RNA suitable for use in downstream applications – Purified RNA 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 analysis requiring the use of intact RNA.
- Multiple kit sizes available – This kit is available in both 50 prep and 100 prep sizes
All prices shown are exclusive of VAT