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| Product Name | Ethidium Bromide Solution (10 mg/mL), Molecular Biology Grade |
| Catalog No. | NATR-HMM-0140 |
| Description | A ready-to-use 10 mg/mL ethidium bromide solution in nuclease-free water for the fluorescent staining of nucleic acids in agarose and polyacrylamide gels. Ethidium bromide is the classic, widely used intercalating dye for DNA and RNA visualization in molecular biology. |
| Intended Use | Visualization of double-stranded DNA and RNA in agarose and polyacrylamide gels under ultraviolet (UV) transillumination for analytical and preparative electrophoresis, with applications in PCR analysis, restriction mapping, cloning verification, and nucleic acid quantification estimation. |
| Principle / Technology | Ethidium bromide (3,8-diamino-5-ethyl-6-phenylphenanthridinium bromide) is a planar aromatic cation that intercalates between adjacent base pairs of double-stranded nucleic acids. Upon intercalation, the dye's fluorescence quantum yield increases approximately 20- to 30-fold. When excited by UV radiation (peak excitation ~300 nm with secondary peak at ~520 nm), the intercalated dye emits orange-red fluorescence (peak emission ~605 nm), enabling sensitive detection of nucleic acid bands. |
| Detection Method | UV transillumination at 254, 302, or 365 nm; CCD or Polaroid photographic documentation; densitometry for semi-quantitative band intensity measurement; compatible with blue-light transilluminators at reduced sensitivity |
| Sample Type | Double-stranded DNA and RNA separated on agarose or polyacrylamide gels; applicable for both analytical and preparative electrophoresis prior to gel extraction; also usable for solution-based fluorescence quantification with appropriate spectrophotometer |
| Performance Range / Specifications | Concentration: 10 mg/mL in nuclease-free water; typical usage: 0.5 μg/mL final in gel or running buffer (1:20,000 dilution); detection sensitivity: ~1–5 ng DNA per band; binding: 1 molecule per 2–3 bp (at saturating concentration); linear dynamic range for densitometry: 10–500 ng per band |
| Sensitivity / LOD | Detects approximately 1–5 ng of double-stranded DNA per band under standard UV transillumination; approximately 50–100 ng for single-stranded RNA under denaturing conditions; detection limit depends on UV transilluminator intensity and imaging system sensitivity |
| Specificity | Intercalates into double-stranded nucleic acids with high affinity (Ka ~10^5–10^6 M^-1); binds preferentially to double-stranded over single-stranded nucleic acids; exhibits low binding to proteins and other cellular macromolecules under standard electrophoresis conditions |
| Reaction Conditions / Protocol | For gel staining: add ethidium bromide to molten agarose (cooled to 50–60°C) at 0.5 μg/mL final concentration before pouring; alternatively, post-stain gel in electrophoresis buffer containing 0.5 μg/mL ethidium bromide for 15–30 minutes followed by destaining in water or buffer for 10–30 minutes; for running buffer staining: add dye to cathode buffer reservoir at 0.5 μg/mL |
| Components / Formulation | Ethidium bromide (≥95% purity by HPLC), 10 mg/mL in nuclease-free, distilled water, supplied in an amber or light-protective bottle |
| Storage Conditions | Room temperature (15–25°C) in tightly sealed, light-protective amber bottle; avoid exposure to strong light which can cause photobleaching; do not freeze; stable for years under recommended conditions |
| Shelf Life | 48 months from date of manufacture in unopened, properly stored container |
| Package Specifications | 1 mL, 5 mL, and 10 mL dropper bottles; 50 mL and 100 mL bulk laboratory bottles |
| Product Form | Ready-to-use dark red liquid solution (10 mg/mL) |
| Quality Control | Each lot tested: DNA detection sensitivity: 5 ng λ/HindIII fragment visible; chemical purity ≥95% by HPLC; absorption spectrum: λmax at 285 nm and 480 nm with correct absorbance ratios; no detectable DNase/RNase activity; PCR compatibility verified (no inhibition at residual dye levels after gel extraction) |
| Key Features | Extremely high sensitivity for routine DNA gel staining; cost-effective compared to fluorescent alternatives; rapid staining with pre-cast or post-stain protocols; chemically stable with indefinite shelf life when properly stored; compatible with all standard UV gel documentation systems; extensive published protocols and safety guidelines available |
For research use only, not for clinical use.
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