A260 Absorbance to Concentration Calculator

Convert UV absorbance at 260 nm (A260) to concentration for dsDNA, ssDNA, or RNA. Tailored for cuvette spectrophotometer measurements with adjustable dilution factors and path lengths.

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Measurement Inputs

Input absorbance, nucleic acid type, and dilutions.

Nucleic Acid Concentration

25.0 μg/mL

* Equal to 25.0 ng/μL.

Extinction Constant 50 μg/mL per OD
Total Yield 2.5 μg (2500 ng)
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Expert Tip

Ensure your A260 reading falls within the spectrophotometer's linear range (typically 0.1 to 1.0 OD). If the reading is too high, dilute the sample and adjust the dilution factor parameter accordingly.

Methodology & Equations

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Beer-Lambert Law Application

Absorbance spectroscopy relates light extinction to substance concentration:

Conc (μg/mL) = (A260 * Factor * Dilution) / Path_Length
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Standard Conversion Constants

At 260 nm wavelength, an optical density (OD) of 1.0 corresponds to standard mass concentrations:

- dsDNA: 50 μg/mL (or ng/μL)
- ssDNA: 33 μg/mL (or ng/μL)
- RNA: 40 μg/mL (or ng/μL)

How to Convert A260 Absorbance to Concentration

Quantify nucleic acids using UV-spectrophotometry at 260 nm:

1

Record A260 Value

Measure the absorbance of your sample at 260 nm.

2

Select standard constants

Factor = 50 µg/mL for double-stranded DNA, 33 µg/mL for single-stranded DNA, or 40 µg/mL for RNA.

3

Calculate Concentration

Multiply absorbance by the nucleic acid constant and your dilution factor: Concentration = A260 × Constant × Dilution Factor.

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