The RNA BR Assay Kit is a fast, sensitive, and effective quantitative RNA fluorescence identification technique. The kit includes fluorescence detection reagents, buffers, and RNA standards. It has superior linearity between 20 ng and 1000 ng for RNA specimens, enabling precise quantification of initial sample concentrations spanning from 1 ng/L to 1000 ng/L. This kit is strongly selective for total RNA, rRNA, and mRNA, and is unaffected by dsDNA. Meantime, the most common contaminants, such as salt, free nucleotides, proteins, solvents, and detergents, are tolerated by the assay. The assay can be conducted at room temperature. Along with the buffer, the fluorescence detection reagent first should be diluted in a working solution before the Standard or RNA specimen for detection is added by the Qubit fluorometer. All elements of the kit are subjected to a strict quality check procedure.
Workflow:
CD RNA BR Assay Kit Workflow
Storage:
The Kit can be stored at 2-8℃ for 6 months. And protect it away from light and avoid repeated freeze-thaw cycles.
Components:
Specifications:
| Sample amount | 1ng/µL-1000 ng/µL |
| Features | High sensitivity: Enable accurate quantification for initial sample concentrations from 1ng/µL to 1000 ng/µL. High specificity: Highly selective for total RNA, rRNA, mRNA, not affected by dsDNA and most conventional contaminations, including salt, free nucleotides, proteins, solvents, and detergents are well-tolerated. Easy operation: Dilute the fluorescence detection reagent with its buffer into a working solution, and then add the Standard or RNA sample for detection by a Qubit fluorometer. The assay can be performed at room temperature. |
| Sample type | RNA |
Note: This protocol is only suitable for Qubit® 2.0, Qubit® 3.0 and Qubit® 4.0 fluorimeters.
(1) All the kit components to room temperature before use.
(2)Prepare sufficient 0.5 ml PCR tubes to accommodate all samples and standards.
Note: Only 0.5ml PCR tubes are suitable for detection.
(3) Label the lid of each tube. DO NOT label on the side wall, in order to avoid any possible interference in fluorescence signal acquisition.
(4) Prepare fresh working solution of CD RNA BR Reagent, by diluting it in CD RNA BR Buffer according to a ratio of 1:200. DO NOT use glass containers for the preparation of working solution.
Note: A sufficient amount of working solution should be prepared to accommodate all samples and standards. For example, to accommodate 7 RNA samples and 2 standards, it is needed to prepare 2 ml of working solution by adding 10 μl of CD RNA BR Reagent to 1990 μl of CD RNA BR Buffer. Mix thoroughly by vortexing.
(5) Prepare standards. Load 190 μl of working solution to each tube used for standards, then carefully add exactly 10 μl of Standard #1 and Standard #2 to the corresponding tube, respectively. Gently vortex for 2-3 sec to avoid bubbles. Make sure the exact pipetting of 10 μl.
(6) Prepare samples. For each tube used for samples, add 180 μl - 199 μl of working solution, and then carefully add 1 μl - 20 μl of RNA sample. Make sure the final volume in each tube is 200 μl. Gently vortex for 2-3 sec to avoid bubbles.
(7) Incubate at room temperature for 2 min. Protect from light.
(8) Load the sample tubes into a Qubit® Fluorometer and test the sample concentration by performing the RNA Broad Range Detection Program.
Application Notes
(1) Protect from light during storage to avoid quenching of fluorescent dyes.
(2) For all reagent and standards in this kit, please mix thoroughly before use by gently inverting tube. Centrifuge for 1-2 sec to collect the reagents to the bottom of tube.
(3) Carefully pipetting of exact volume is critical to ensure accurate quantification. Please use calibrated pipettes for the assay.
(4) Please perform the assay at room temperature. All the kit components to room temperature before use. DO NOT hold the tubes for assay in hands for a long time.
(5) The working solution in Step 4 should be prepared fleshly and used within 3 hours, to avoid fluorescence quenching.
(6) To avoid degradation of RNA standards, use RNA-free consumables for the experiment and place the standards at 2-8 °C after the end of the experiment.
