Abstract
Any experiment requiring manipulation of a nucleic acid most likely also requires its accurate quantification to ensure optimal and reproducible results. This chapter shows the relationship between nucleic acid concentration and absorption of ultraviolet light. That relationship can be used to determine the concentration of double-stranded DNA, single-stranded DNA, synthetic oligonucleotides, and RNA. Multiple methods are presented for expressing nucleic acid concentration with different units (μg/mL, millimolar, etc.). An extinction coefficient is used to calculate nucleic acid concentration expressed as a millimolar amount. Fluorescent dyes can be used to extrapolate the concentration of small amounts of nucleic acid by utilizing a standard curve. This chapter also describes how to estimate nucleic acid concentration by staining following electrophoresis as well as how to calculate a nucleic acid's limit of quantitation and limit of detection.
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