Description
dsDNA broad-range assay kit
GenieQuant dsDNA BR Assay Kit is a fluorometric assay for selective quantification of double-stranded DNA across a broad, high concentration range. It is suited to routine preps such as minipreps and genomic DNA, and its dsDNA-specific dye gives accurate readings unaffected by RNA, nucleotides or protein.
The kit contains a fluorescent dye, buffer and dsDNA standards. It is linear from 4 to 2,000 ng of dsDNA, allowing accurate quantification of sample concentrations from 0.2 ng/μl to 2,000 ng/μl. It is highly selective for dsDNA over RNA and tolerates common contaminants such as RNA, salts, free nucleotides, proteins, solvents and detergents. Simply dilute the dye 1:200 in buffer, add the sample and read on a Qubit Fluorometer after 2 min at room temperature. For research use only.
Key Features
| Precise and sensitive | Accurately quantifies 4–2,000 ng of dsDNA. | |
| High specificity | Detects dsDNA selectively and tolerates common contaminants. | |
| Rapid dye binding | Signal reaches saturation within 2 minutes. | |
| Simple workflow | One working solution, performed entirely at room temperature. |
Specifications
| Method | Fluorescent dsDNA-binding dye |
| Linear range | 4–2,000 ng dsDNA |
| Sample concentration range | 0.2–2,000 ng/μl |
| Sample volume | 1–20 μl (200 μl final assay volume) |
| Incubation | 2 min at room temperature |
| Instrument | Qubit Fluorometer (dsDNA Broad Range program) |
| Shelf life | 2 years |
Components
| Components | 100 assays | 500 assays |
| GenieQuant dsDNA BR Reagent (200× in DMSO) | 250 μl | 1.25 ml |
| GenieQuant dsDNA BR Buffer | 50 ml | 250 ml |
| GenieQuant dsDNA BR Standard #1 (0 ng/μl in TE buffer) | 1 ml | 5 ml |
| GenieQuant dsDNA BR Standard #2 (100 ng/μl in TE buffer) | 1 ml | 5 × 1 ml |
For long-term storage, keep Standard #2 at -30 ~ -15°C.
Storage
Store at 2 ~ 8°C.
This protocol is for the Qubit Fluorometer.
- Equilibrate
- Bring all components to room temperature before use.
- Prepare tubes
- Prepare enough 0.5 ml PCR tubes for all samples and standards. Use only 0.5 ml PCR tubes.
- Label the lid of each tube, not the side, as side labels can interfere with the reading.
- Prepare the working solution
- Freshly dilute GenieQuant dsDNA BR Reagent 1:200 in GenieQuant dsDNA BR Buffer. Do not use a glass container.
- Make enough for all standards and samples at 200 μl per tube. For example, for 7 samples plus 2 standards (9 tubes), mix 10 μl of reagent with 1,990 μl of buffer to give 2 ml.
- Prepare the standards
- Add 190 μl of working solution to each standard tube, then add 10 μl of Standard #1 or Standard #2.
- Prepare the samples
- Add 180–199 μl of working solution to each sample tube, then add 1–20 μl of DNA sample, for a final volume of 200 μl.
- For 1–2 μl samples, use a 2 μl pipette for accuracy.
- Mix
- Vortex gently for 2–3 sec, then centrifuge briefly to collect the liquid and remove bubbles.
- Incubate and read
- Incubate all tubes at room temperature for 2 min, protected from light.
- Measure using the dsDNA Broad Range program, following the Qubit Fluorometer instructions.
Q1: What concentration range can the kit measure?
- The assay is linear from 4 to 2,000 ng of dsDNA, covering sample concentrations from 0.2 ng/μl to 2,000 ng/μl with 1–20 μl of sample.
Q2: Will RNA or other contaminants affect my reading?
- The dye is highly selective for dsDNA over RNA and tolerates common contaminants such as RNA, salts, free nucleotides, proteins, solvents and detergents.
Q3: Which tubes and instrument should I use?
- Use only 0.5 ml PCR tubes, labelled on the lid, and read them on a Qubit Fluorometer using the dsDNA Broad Range program.
Q4: How soon should I read the samples?
- Incubate for 2 min at room temperature, protected from light, and complete the measurement within 3 h of adding the samples.
