2.4.2 Nucleic Acid Quantification

Nucleic acids absorb UV light of 250 to 270 nm wavelength, with a maximum at 260 nm. The reading at 260 nm allows calculation of the concentration of nucleic acid in the sample. An OD of 1 corresponds to approximately 50 µg/ml for double-stranded DNA and 40 µg/ml for single-stranded DNA and RNA. The ratio between the readings at 260 nm and 280 nm (A260280 ratio) provides an estimate for the purity of the nucleic acid. Pure preparations of DNA and RNA have ratios of 1.8 to 1.95, respectively. DNA producing a ratio below 1.6 should be re-purified3.

© 2001 Alexander Binder