Comparison of Fluorescence-Based Quantitation with UV Absorbance Measurements

Detection and quantitation of nucleic acids are vital to many biological studies but the methods commonly used can be unreliable and inaccurate. UV absorbance measurements are not selective and cannot distinguish DNA, RNA, or protein and walues are easily affected by other contaminants and variations in base composition. In addition, the sensitivity of spectrophotometry is often inadequate, prohibiting quantitation of DNA and RNA at low concentrations. This application note compares the two quantitation methods using the Qubit® Fluorometer and a microvolume UV spectrophotometer. The Qubit® fluorometric quantitation provided a more selective, sensitive, and accurate method for quantitating nucleic acids than UV absorbance measurements.