How to calculate od optical density for isolation of genomic dna experiment?
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DNA yield can be assessed using various methods including absorbance (optical density), agarose gel electrophoresis, or use of fluorescent DNA-binding dyes. All three methods are convenient, but have varying requirements in terms of equipment needed, ease of use, and calculations to consider.
Absorbance Methods
The most common technique to determine DNA yield and purity is measurement of absorbance. Although it could be argued that fluorescence measurement is easier, absorbance measurement is simple, and requires commonly available laboratory equipment. All that is needed for the absorbance method is a spectrophotometer equipped with a UV lamp, UV-transparent cuvettes (depending on the instrument) and a solution of purified DNA. Absorbance readings are performed at 260nm (A260) where DNA absorbs light most strongly, and the number generated allows one to estimate the concentration of the solution. To ensure the numbers are useful, the A260 reading should be within the instrument's linear range.
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