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Updated: Jul 5, 2026

NanoDrop Microvolume Quantitation of Nucleic Acids

NanoDrop Microvolume Quantitation of Nucleic Acids

Published on: November 22, 2010

Quantitation of nucleic acids with absorption spectroscopy.

¹Motorola Corporation, Tempe, Arizona, USA.

Current Protocols in Protein Science

|April 23, 2008

Summary

This protocol details measuring nucleic acid purity and concentration using absorbance at different wavelengths. The ratio of absorbance at 260 nm and 280 nm indicates sample purity.

Area of Science:

Biochemistry
Molecular Biology
Analytical Chemistry

Background:

Accurate quantification and purity assessment of nucleic acids are crucial for molecular biology applications.
Spectrophotometric analysis is a common method for evaluating nucleic acid samples.

Purpose of the Study:

To describe a protocol for measuring nucleic acid absorption at multiple wavelengths.
To establish methods for assessing nucleic acid purity and concentration.

Main Methods:

Measuring absorbance of nucleic acid samples at various wavelengths.
Utilizing A(260) measurements for quantitative analysis of microgram quantities.
Calculating the ratio of absorbance at 260 nm and 280 nm (A260/A280) to assess purity.

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Main Results:

Absorbance at 260 nm (A260) provides quantitative data for pure nucleic acid preparations.
Absorbance readings alone cannot differentiate between DNA and RNA.
The A260/A280 ratio serves as a reliable indicator of nucleic acid purity.

Conclusions:

The described protocol enables accurate determination of nucleic acid concentration and purity.
Spectrophotometric analysis, particularly the A260/A280 ratio, is essential for quality control in molecular biology.