Abstract
This lab will cover ultraviolet-visible (UV-Vis) spectroscopy to simultaneously assay the quality and quantity of extracted DNA. This experiment will use a DNA sample of at least two micrograms made visible in a traditional one milliliter sample using a conventional instrument or two nanograms per microliter using a small volume NanoDrop instrument. The concentration of the DNA-containing sample can be computed by dividing the measured absorbance by the path length and absorptivity coefficient of 50 μg/mL for double-stranded DNA and 33 μg/mL for single-stranded DNA. The quality of the recovered DNA is determined by the ratio of the absorbances at 260 nm to 280 nm for each sample. A high quality DNA sample has a 260 nm / 280 nm ratio of 1.7 to 2.0. The method is non-destructive and the DNA may be recovered from the cuvette by pipetting. The quality and quantity of the recovered DNA will be reported.
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