Quantifying and Storing RNA.

Abstract

Quantifying RNA is an important and necessary step before most RNA analysis techniques. Methods for quantifying RNA can be classified into two categories: ultraviolet (UV) spectrophotometric methods, which are based on the absorption spectra of the purine and pyrimidine bases; and fluorescent dye-based methods, which measure the fluorescence intensity of dyes that selectively fluoresce when bound to nucleic acids. If the RNA sample is pure (i.e., without significant amounts of contaminants such as proteins, phenol, agarose, or other nucleic acids), UV spectrophotometric measurement of the amount of UV irradiation absorbed by the bases is simple and accurate. However, if the sample contains significant quantities of impurities or if the concentration of RNA is very low, it is better to use fluorescent dye-based methods. An overview of spectrophotometric and fluorescent dye-based RNA quantification methods is given here, as are several options for storing purified RNA preparations. Proper storage of RNA samples is important; it can help minimize RNase contamination and consequent sample degradation.