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| Product Name | Qubit dsDNA High-Sensitivity Quantification Kit |
| Catalog No. | NATR-HMM-0127 |
| Description | A highly sensitive fluorescence-based assay kit for the accurate quantification of double-stranded DNA (dsDNA) in dilute samples. The kit employs a target-selective fluorogenic dye that emits fluorescence only when bound to dsDNA, providing superior specificity compared to UV absorbance-based methods. |
| Intended Use | Precise quantitation of dsDNA in dilute samples for next-generation sequencing library preparation, qPCR template normalization, and any application requiring accurate DNA concentration determination independent of sample contaminants. |
| Principle / Technology | A proprietary dsDNA-selective fluorescent dye exhibits minimal intrinsic fluorescence in the unbound state but produces a strong fluorescence signal upon intercalation into double-stranded DNA. The fluorescence intensity is directly proportional to dsDNA concentration over the calibrated range. Unlike UV A260 measurement, the assay is unaffected by free nucleotides, ssDNA, RNA, protein, salts, solvents, or detergents. |
| Detection Method | Fluorescence measurement using a benchtop fluorometer (excitation: 470–485 nm, emission: 525–535 nm); two-point standard calibration for concentration determination |
| Sample Type | Purified dsDNA from any source including genomic DNA, plasmid DNA, PCR products, restriction fragments, and NGS library preparations; compatible with common DNA purification elution buffers (TE, EB, nuclease-free water) |
| Performance Range / Specifications | Detection range: 0.2–100 ng per assay tube (0.01–5 ng/μL in a 20 μL sample volume); linear correlation: R² ≥0.995 over full dynamic range; accuracy: ±10% of nominal concentration for samples within range; precision: CV ≤5% for replicate measurements |
| Sensitivity / LOD | Lower limit of detection: 0.01 ng/μL (200 pg in 20 μL assay); reliably distinguishes 2-fold concentration differences within the assay range |
| Specificity | Highly selective for double-stranded DNA; negligible signal from RNA (even at 10× excess), single-stranded DNA, proteins, free nucleotides, salts, phenol, ethanol, chloroform, detergents (≤0.1% SDS, ≤1% Triton X-100), agarose, and common DNA extraction contaminants |
| Reaction Conditions / Protocol | Prepare working solution by diluting dye reagent 1:200 in buffer, add 190 μL working solution to each assay tube, add 10 μL of each standard (0 and 100 ng/μL) and sample, vortex briefly, incubate at room temperature for 2 minutes, read fluorescence on fluorometer; calculation is automated by instrument software |
| Components / Formulation | Qubit dsDNA HS reagent (dsDNA-specific fluorescent dye in DMSO), Qubit dsDNA HS buffer, dsDNA standards (0 ng/μL and 100 ng/μL in TE buffer with preservative), thin-walled 0.5 mL assay tubes (500 tubes included) |
| Storage Conditions | 2–8°C for reagent and buffer (allow to warm to room temperature before use); standards stored at 2–8°C; protect reagent from light; working solution stable for 2–3 hours at room temperature protected from light |
| Shelf Life | 12 months from date of manufacture at recommended storage |
| Package Specifications | 100 assays and 500 assays; includes enzyme, buffer, standards, and compatible assay tubes |
| Product Form | Liquid dye reagent concentrate, buffer, and DNA standards |
| Quality Control | Each lot validated: standard curve R² ≥0.995; detection of 0.01–100 ng DNA per assay; no cross-reactivity with 100 ng/μL RNA; accuracy ±10% against NIST-traceable DNA standards; precision CV ≤5%; functionality with common extraction kit eluents verified |
| Key Features | dsDNA-specific dye eliminates overestimation from RNA and ssDNA contamination; 1000× more sensitive than UV A260 measurement; 2-minute incubation for rapid workflow; minimal interference from common extraction contaminants; narrow-range kits available for ultra-low concentration samples |
For research use only, not for clinical use.
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