The RNA Subcellular Isolation Kit provides an efficient method to isolate nuclear and cytoplasmic RNA without cross-contamination. This method can be used to isolate RNA sequences as small as 75 nucleotides (e.g. snoRNA, mRNA, and lncRNA). RNA molecules exist at different levels of maturation and processing within a cell at any given time. Such heterogeneity can bias downstream analysis and interpretation of data. By isolating RNA from specific subcellular fractions, the accuracy of gene expression analysis can be greatly improved. Subcellular isolation enhances the ability to detect low abundance transcripts and aids in the ability to study RNA processing dynamics by reducing the background from other intronic or mature RNAs. Additionally, subcellular isolation helps to localize long non-coding RNAs (lncRNAs) to their site of action, which can provide important insights into their functional role.
The RNA Subcellular Isolation Kit contains enough reagents to perform 15 nuclear and 15 cytoplasmic purifications from 2-4 million cultured cells or 15 mg of tissue per reaction. Simply lyse your samples, then separate the nuclear and cytoplasmic RNA by centrifugation. Each RNA fraction is loaded onto individual spin columns and washed to remove any proteins. Finally, the purified RNA is eluted from the column and quantified. The assay can also be used to isolate total RNA.
