Nucleic acids are quantified at 260 nm — deep UV, where disposable plastic and optical glass are both opaque. If you are measuring DNA or RNA by absorbance, a quartz cuvette is not optional. This guide covers the measurement, how to read the two purity ratios that tell you whether your sample is actually clean, and how to pick path length and volume.
The absorbance maximum of the nucleotide bases sits at 260 nm. Measuring there gives concentration directly from Beer-Lambert, with no reagents and no consumption of the sample beyond the measurement itself.
Material cut-offs make the choice for you: polystyrene and optical glass both stop transmitting around 340 nm. At 260 nm they absorb essentially everything, so a plastic cuvette returns noise, not data. JGS1 fused silica transmits from 200 nm, placing 260 nm well inside its usable window.
At a 10 mm path length, an absorbance of 1.0 corresponds to:
| Nucleic acid | Concentration at A260 = 1.0 (10 mm) |
|---|---|
| dsDNA | 50 µg/mL |
| ssDNA | 33 µg/mL |
| RNA | 40 µg/mL |
| Oligonucleotides | ~33 µg/mL (varies with base composition) |
Values assume a 10 mm path. For other path lengths, divide by the path in cm. The path length calculator handles the arithmetic.
| Ratio | Clean value | Low value indicates |
|---|---|---|
| A260 / A280 | ≈1.8 (DNA), ≈2.0 (RNA) | Protein or phenol contamination |
| A260 / A230 | 2.0 – 2.2 | Salts, carbohydrates, phenol, guanidine, TRIzol carry-over |
Both ratios only mean something if the absorbance values sit in the linear range (roughly 0.1–1.0 AU). A ratio calculated from a 0.02 AU reading is noise dressed up as data — dilute less or use a longer path.
Extraction yields are small and getting smaller. The standard 10 mm macro cell wants ~3.5 mL, which no modern extraction protocol produces.
Pedestal instruments measure 1–2 µL and are convenient, but they cap the measurable concentration at the top end and are sensitive to how the droplet is formed and whether the pedestal is perfectly clean. A quartz cuvette is more reproducible, is easy to validate, and costs far less per sample over its life. We compared both in depth: Cuvette vs NanoDrop.
Related: Cleaning Protocol · UV-Vis Troubleshooting · Solvent Compatibility · Path Length Guide · Path Length Calculator · Cuvette Care Guide
Quartz 10 mm cells for A260 readings (UV); micro or ultra-micro when sample is limited. Protein quantification.
A260 is in the UV (about 260 nm), below glass transmission; quartz (190–2500 nm) is required. UV cutoff.
Yes — ultra-micro down to about 35 µL preserves precious nucleic-acid prep. Ultra-micro.
10 mm (1 cm) matches the standard conversion (A260 = 1 is about 50 µg/mL dsDNA). Path length guide.
Use RNase-free rinses; avoid cross-contamination. Cleaning protocol.