Description
RNA high-sensitivity assay kit
GenieQuant RNA HS Assay Kit is a fluorometric assay for highly sensitive, selective quantification of RNA at low concentrations. It is ideal for limited or low-input samples and library QC, and its RNA-specific dye gives accurate readings without interference from DNA, free nucleotides or protein.
The kit contains a fluorescent detection reagent, buffer and RNA standards. It is highly selective for RNA over double-stranded DNA and is linear over 4–200 ng RNA, allowing accurate quantification of total RNA, rRNA and mRNA from 200 pg/μl to 200 ng/μl. It tolerates common contaminants such as salts, free nucleotides, proteins, solvents and detergents. The assay runs at room temperature: dilute the reagent 1:200 in buffer, add the sample, incubate for 2 min and read on a Qubit 2.0, 3.0 or 4.0 fluorometer. For research use only.
Key Features
| High sensitivity | Accurately quantifies total RNA, rRNA and mRNA from 200 pg/μl to 200 ng/μl. | |
| High specificity | Highly selective for RNA, unaffected by DNA, and tolerant of salts, free nucleotides, proteins, solvents and detergents. | |
| Easy to use | Mix the working solution with sample and read after 2 min at room temperature. |
Specifications
| Method | Fluorometric RNA quantification |
| Detection range | 200 pg/μl - 200 ng/μl (linear over 4–200 ng RNA) |
| Sample types | Total RNA, rRNA, mRNA |
| Sample volume | 1–20 μl in a 200 μl assay |
| Instrument | Qubit 2.0, 3.0 and 4.0 fluorometers |
| Shelf life | 2 years |
Components
| Components | 100 assays | 500 assays |
| GenieQuant RNA HS Reagent (200× in DMSO) | 250 μl | 1.25 ml |
| GenieQuant RNA HS Buffer | 50 ml | 250 ml |
| GenieQuant RNA HS Standard #1 (0 ng/μl in TE buffer) | 1 ml | 5 ml |
| GenieQuant RNA HS Standard #2 (10 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 only.
- Equilibrate
- Bring all components to room temperature before use.
- Prepare tubes
- Use only 0.5 ml PCR tubes, one for each sample and standard.
- Label the tube lids, not the sides, as side labels can interfere with the reading.
- Prepare the working solution
- Freshly dilute the GenieQuant RNA HS Reagent 1:200 in GenieQuant RNA HS Buffer. Do not mix in a glass container.
- Make enough for all samples and both standards at 200 μl per tube. For example, for 7 samples plus 2 standards (9 tubes), mix 10 μl reagent with 1990 μl buffer to make 2 ml.
- Prepare the standards
- Add 190 μl working solution to each standard tube, then 10 μl of 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 1–20 μl RNA sample, for 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
- Following the fluorometer instructions, select the RNA high-sensitivity program and measure the concentration.
Q1: What concentration range can the kit measure?
- It quantifies total RNA, rRNA and mRNA from 200 pg/μl to 200 ng/μl, with linear response over 4–200 ng RNA in the assay.
Q2: Does DNA in the sample interfere?
- The assay is highly selective for RNA over double-stranded DNA and tolerates common contaminants such as salts, free nucleotides, proteins, solvents and detergents.
Q3: Which instruments and tubes are compatible?
- The protocol is for Qubit 2.0, 3.0 and 4.0 fluorometers, using 0.5 ml PCR tubes only.
Q4: How long is the working solution stable?
- Complete measurements within 3 h of preparing the working solution, as fluorescence quenching can otherwise give inaccurate results.
