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| Product Name | PicoGreen dsDNA Quantitation Reagent |
| Catalog No. | NATR-HMM-0141 |
| Description | An ultrasensitive fluorescent nucleic acid stain designed for the precise quantitation of double-stranded DNA in solution. PicoGreen dye offers exceptional sensitivity with a linear detection range spanning four orders of magnitude using standard fluorescence microplate readers or fluorometers. |
| Intended Use | Accurate low-concentration dsDNA measurement for next-generation sequencing library quantification, forensic DNA analysis, quantitative PCR template normalization, viral DNA titer determination, and applications requiring precise DNA measurement at picogram-level sensitivity. |
| Principle / Technology | PicoGreen reagent is an asymmetric cyanine dye that exhibits minimal intrinsic fluorescence in the unbound state but undergoes a >1,000-fold fluorescence enhancement upon specific binding to double-stranded DNA. The dye intercalates between base pairs with high affinity, producing a fluorescence signal directly proportional to dsDNA concentration over four orders of magnitude. The assay is highly selective for dsDNA over single-stranded DNA, RNA, proteins, and free nucleotides. |
| Detection Method | Fluorescence measurement using a standard fluorescence microplate reader, fluorometer, or fluorescence spectrophotometer (excitation: ~480 nm, emission: ~520 nm); compatible with black 96-well and 384-well plates for high-throughput quantification |
| Sample Type | Purified dsDNA from any biological source or synthetic origin; samples may contain common contaminants including salts, proteins, detergents, and organic solvents without interfering with quantitation; compatible with samples eluted in TE, EB, or nuclease-free water |
| Performance Range / Specifications | Linear detection range: 25 pg/mL to 1 μg/mL dsDNA when using the standard assay protocol (four orders of magnitude); accuracy: within 5% of known concentration across linear range; assay time: 2–5 minutes incubation at room temperature; sample volume: 1–100 μL (with standard or microplate protocols) detectable in 200 μL final assay volume |
| Sensitivity / LOD | Detects as little as 25 pg/mL dsDNA (2.5 pg in a 100 μL sample, equivalent to ~25 pg in a standard 200 μL assay); lower limit of detection approximately 100-fold more sensitive than Hoechst 33258-based assays and >10,000-fold more sensitive than A260 absorbance |
| Specificity | Highly selective for double-stranded DNA; negligible fluorescence enhancement with single-stranded DNA, RNA, proteins, detergents, and common laboratory chemicals at typical sample concentrations; RNA at 1 μg/mL generates <0.5% of the signal of an equal mass of dsDNA |
| Reaction Conditions / Protocol | Prepare aqueous working solution by diluting PicoGreen reagent 1:200 in TE buffer (10 mM Tris-HCl pH 7.5, 1 mM EDTA), protect from light; in a microplate well or cuvette, add 100 μL sample and 100 μL diluted working reagent (final dilution 1:400), mix, incubate 2–5 minutes protected from light, measure fluorescence at 480/520 nm; prepare dsDNA standard curve (0–1 μg/mL) concurrently with each assay for accurate quantitation |
| Components / Formulation | PicoGreen dsDNA reagent (proprietary cyanine dye in anhydrous DMSO, supplied as 1 mL concentrate), 20× TE buffer concentrate, lambda DNA standard (100 μg/mL), detailed protocol |
| Storage Conditions | Reagent: –20°C protected from light and moisture (desiccated); working solution: prepare fresh and use within 2–3 hours; dye concentrate is stable through multiple freeze-thaw cycles if properly stored; TE buffer and DNA standard: 2–8°C or –20°C |
| Shelf Life | 12 months at –20°C from date of manufacture in unopened vial; after opening, protect from light and moisture |
| Package Specifications | 10 mL (sufficient for 2,000 assays in 200 μL format) and 100 mL bulk (20,000 assays); high-throughput packaging available |
| Product Form | Liquid dye concentrate in DMSO; ready to dilute in aqueous buffer for use |
| Quality Control | Each lot tested: 1 μg/mL calf-thymus dsDNA standard produces fluorescence ≥100× blank; linear calibration curve R² ≥0.995 from 0–1 μg/mL; dsDNA selectivity verified: ssDNA signal <2% of dsDNA at equal mass; RNA signal ≤0.5% of dsDNA; lot-to-lot fluorescence intensity within ±10%; DNase/RNase-free by functional assay |
| Key Features | >10,000-fold more sensitive than A260 absorbance; four-log linear dynamic range eliminates serial dilution; dsDNA-selective dye eliminates RNA and ssDNA interference; compatible with 96- and 384-well fluorescence plate readers for high-throughput; room-temperature stable working solution for batch processing |
For research use only, not for clinical use.
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