핵산은 260 nm 심자외선에서 정량합니다. 일회용 플라스틱과 광학 유리 모두 이 파장을 투과하지 못합니다. 흡광도로 DNA나 RNA를 측정한다면 석영 큐벳은 선택이 아닌 필수입니다. 본 가이드는 측정법, 두 가지 순도 비율 해석, 광로와 용량 선택을 다룹니다.
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
A260 측정(UV)용 석영 10 mm 셀. 시료가 제한되면 마이크로 또는 울트라마이크로입니다. 단백질 정량.
A260은 UV(약 260 nm)에 있으며 유리 투과 영역 미만입니다. 석영(190–2500 nm)이 필요합니다. UV 컷오프.
예 — 울트라마이크로는 약 35 µL까지로 귀중한 핵산 조제를 보존합니다. 울트라마이크로.
10 mm(1 cm)는 표준 환산(A260 = 1은 이중가닥 DNA 약 50 µg/mL)과 일치합니다. 광경로 길이 가이드.
RNase 프리 헹굼을 사용하고 교차 오염을 피하세요. 세척 프로토콜.