How can one quantify the amount of nucleic acid that is present in a sample and
determine its purity?
What will be an ideal response
Both DNA and RNA absorb ultraviolet light at a wavelength of 260 nm in direct proportion to their concentration
in a sample. If a 1 cm path length is used, the extinction coefficient can be used to quantify the nucleic acid present.
The factor for DNA is 50 ug/mL, and the factor for RNA is 40 ug/mL. For example, the absorbance of a DNA
sample at 260 nm must be multiplied by 50 and the sample dilution factor to calculate how many ug of DNA are
present in the sample. Purity usually means freedom from proteins which absorb well at 280 nm. If a sample is read
at both 260 and 280 nm and the nucleic acid is pure, the ratio should be 1.8 for DNA and 2.0 for RNA.
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