Description
RNA broad-range assay kit
GenieQuant RNA BR Assay Kit is a fluorometric assay for selective quantification of RNA across a broad, high concentration range, suited to routine RNA extractions. The RNA-specific dye gives accurate measurement unaffected by DNA, free nucleotides or protein.
The kit contains a fluorescent detection reagent, buffer and two RNA standards. It is highly selective for RNA over double-stranded DNA and is linear for 20–1,000 ng RNA in the assay, so total RNA, rRNA and mRNA can be quantified at sample concentrations of 1–1,000 ng/μl. It tolerates common contaminants such as salts, free nucleotides, proteins, solvents and detergents. The assay runs at room temperature: dilute the reagent in buffer to make the working solution, add sample, and read on a Qubit 2.0, 3.0 or 4.0 Fluorometer. For research use only.
Key Features
| Broad range | Accurately quantifies total RNA, rRNA and mRNA from 1 ng/μl to 1,000 ng/μl. | |
| High specificity | Highly selective for RNA and not affected by DNA. | |
| Contaminant tolerant | Resists interference from salts, free nucleotides, proteins, solvents and detergents. | |
| Simple workflow | Room-temperature assay with a 2 min incubation. |
Specifications
| Method | Fluorometric (RNA-selective dye) |
| Quantification range | 1–1,000 ng/μl (linear for 20–1,000 ng RNA) |
| Sample types | Total RNA, rRNA, mRNA |
| Sample volume | 1–20 μl (200 μl final assay volume) |
| Instrument | Qubit 2.0, 3.0 and 4.0 Fluorometers |
| Shelf life | 2 years |
Components
| Components | 100 assays | 500 assays |
| GenieQuant RNA BR Reagent (200X in DMSO) | 250 μl | 1.25 ml |
| GenieQuant RNA BR Buffer | 50 ml | 250 ml |
| GenieQuant RNA BR Standard #1 (0 ng/μl in TE buffer) | 1 ml | 5 ml |
| GenieQuant RNA BR Standard #2 (100 ng/μl in TE buffer) | 4 × 250 μl | 10 × 500 μl |
Storage
Store at 2 ~ 8°C.
This protocol is for Qubit 2.0, 3.0 and 4.0 Fluorometers.
- Prepare
- Equilibrate all components to room temperature.
- Prepare enough 0.5 ml PCR tubes for all samples and standards (use only 0.5 ml PCR tubes).
- Label the tube lids, not the sides, as side labels can interfere with the reading.
- Make the working solution
- Freshly dilute GenieQuant RNA BR Reagent 1:200 in GenieQuant RNA BR Buffer. Do not use a glass container.
- Make enough for all samples plus 2 standards at 200 μl per tube. For example, for 7 samples (9 tubes), mix 10 μl Reagent with 1,990 μl Buffer to give 2 ml.
- Prepare the standards
- Add 190 μl working solution to each standard tube, then add 10 μl Standard #1 or Standard #2. Pipette accurately.
- Vortex gently for 2–3 sec, avoiding bubbles.
- Prepare the samples
- Add 180–199 μl working solution to each sample tube, then add 1–20 μl RNA sample to a final volume of 200 μl.
- Vortex gently for 2–3 sec, avoiding bubbles.
- Incubate
- Incubate all tubes at room temperature for 2 min, protected from light.
- Read
- Select the RNA Broad Range program on the Qubit Fluorometer and measure according to the instrument instructions.
Q1: What concentration range does the kit measure?
- Total RNA, rRNA and mRNA at 1–1,000 ng/μl in the original sample, with a linear response for 20–1,000 ng RNA in the assay.
Q2: Will DNA in my sample affect the result?
- No. The dye is highly selective for RNA over dsDNA, and the assay tolerates salts, free nucleotides, proteins, solvents and detergents.
Q3: How long is the working solution stable?
- Complete measurements within 3 h of preparing the working solution to avoid fluorescence quenching.
Q4: Which instruments and tubes are compatible?
- The protocol is for Qubit 2.0, 3.0 and 4.0 Fluorometers and uses 0.5 ml PCR tubes only.
