Quantification of Nucleic Acids: A260/A280 ratio, Nucleic Acid Concentration, Purity and Molarity

Any experiment requiring manipulation of a nucleic acid most likely also requires its accurate quantification to ensure optimal and reproducible results. The nitrogenous bases positioned along a nucleic acid strand absorb ultraviolet (UV) light at a wavelength of 260 nm;
at this wavelength, light absorption is proportional to nucleic acid concentration. This relationship is so well characterized that UV absorption is used to accurately determine the concentration of nucleic acids in solution.
The relationship between DNA concentration and absorption is linear up to an absorption at 260 nm (A260) of 2. For measuring the absorption of a nucleic acid solution in a spectrophotometer, most molecular biology laboratories will use quartz cuvettes with a width through which the light beam will travel 1 cm.
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