The dsDNA HS Assay Kit is a fluorescence quantitative double-stranded DNA detection technique (dsDNA). It's fast, sensitive, and precise and t includes fluorescence detection reagents, buffers, and dsDNA standards. It has excellent linearity for dsDNA specimens ranging from 0.2 to 100 ng, permitting precise quantification of initial sample concentrations varying from 10 pg/L to 100 ng/L. RNA has no effect on this kit because it only identifies double-stranded DNA. Meanwhile, most common contaminants, such as salt, free nucleotides, proteins, solvents, and detergents, are tolerated by the assay. The assay can be performed at room temperature. The fluorescence detection reagent is diluted in a working solution with its buffer before the Standard or dsDNA specimen is added for detection by a Qubit fluorometer. All Kit parts have undergone a strict quality control process.
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 | 10 pg/µL - 100 ng/µL |
| Features | High sensitivity: Enable accurate quantification for initial sample concentrations from 10 pg/µL to 100 ng/µL. High specificity: Highly selective for double-stranded DNA, not affected by RNA 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 dsDNA sample for detection by a Qubit fluorometer. The assay can be performed at room temperature. |
| Sample type | dsDNA |
Note: This protocol is only suitable for Qubit® 2.0 and Qubit® 3.0 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 dsDNA HS Reagent, by diluting it in CD dsDNA HS 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 dsDNA samples and 2 standards, it is needed to prepare 2 ml of working solution by adding 10 μl of CD dsDNA HS Reagent to 1990 μl of CD dsDNA HS 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, and then carefully add 1 μl - 20 μl of dsDNA 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 dsDNA High Sensitivity Detection Program.
Application Notes
(1) Protect from light during storage and assay 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.
