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Helixyte™ Total DNA Quantification Assay Kit *4-2000 ng Broad Range*
Product key features
- High DNA Specificity: Selectively quantifies all DNA species with minimal interference from RNA or proteins.
- Broad Detection Range: Accurately measures 4–2000 ng of DNA.
- Ready-to-Use Reagents: Pre-formulated 1X working solution minimizes preparation and ensures consistency.
- Contaminant Tolerance: Maintains accuracy in the presence of salts, free nucleotides, detergents, and proteins.
- Fluorescence-Based: Compatible with microplate readers for precise, high-throughput quantification.
Product description
The Helixyte™ Total DNA Quantification Assay Kit offers a streamlined solution for accurately measuring DNA concentrations. With a pre-formulated, ready-to-use working solution, this kit simplifies workflows. Just prepare your sample, mix it with the Helixyte™ Green DNA BR reagent, and measure the DNA concentration using a fluorescence microplate reader.
This assay is optimized for high selectivity toward DNA over RNA, ensuring precise and reproducible quantification. It accommodates a wide range of DNA concentrations and reliably detects 4–2000 ng of DNA. The straightforward protocol involves diluting 1–50 μL of the sample into the provided 1X working solution and recording fluorescence intensity.
The kit is designed to tolerate common contaminants, including salts, free nucleotides, solvents, detergents, and proteins, minimizing interference and maximizing accuracy. Its highly specific formulation ensures superior detection of DNA over RNA and protein, making it ideal for a range of research and diagnostic applications requiring reliable and precise DNA quantitation.
Example protocol
AT A GLANCE
Add test samples (50 µL)
Add Helixyte™ Green DNA BR reagent working solution (50 µL)
Incubate at room temperature for 2 minutes
Monitor fluorescence intensity at Ex/Em = 490/530 nm
Important: The following protocol serves as an example for quantifying total DNA with Helixyte™ Green DNA BR. Ensure all components are brought to room temperature before opening. Please note that no data are available for the mutagenicity or toxicity of Helixyte™ Green DNA BR stain. As this reagent binds to nucleic acids, it should be treated as a potential mutagen and handled with appropriate care. Exercise particular caution when handling the DMSO stock solution, as DMSO is known to facilitate the absorption of organic molecules into tissues.
PREPARATION OF STANDARD SOLUTIONS
https://www.aatbio.com/tools/serial-dilution/17633
PREPARATION OF WORKING SOLUTION
Add 50 μL of Helixyte™ Green DNA BR (Component A) to 2.5 mL of DNA Assay Buffer (Component B) and mix thoroughly. Protect the working solution from light by covering it with foil or storing it in the dark.
Note: We recommend preparing this solution in a plastic container rather than glass, as the dye may adsorb to glass surfaces. For optimal results, use this solution within 2 hours of preparation.
SAMPLE EXPERIMENTAL PROTOCOL
Table 1. Layout of dsDNA standards and test samples in a solid black 96-well microplate. DS=DNA Standards (DS1-DS13, 40 to 0.0098 μg/mL); BL=Blank Control; TS=Test Samples.
BL | BL | DS8 | DS8 |
DS1 | DS1 | DS9 | DS9 |
DS2 | DS2 | DS10 | DS10 |
DS3 | DS3 | DS11 | DS11 |
DS4 | DS4 | DS12 | DS12 |
DS5 | DS5 | DS13 | DS13 |
DS6 | DS6 | TS | TS |
DS7 | DS7 | TS | TS |
Table 2. Reagent composition for each well.
Well | Volume | Reagent |
DS1–DS7 | 50 µL | Serial Dilutions (40 to 0.0098 μg/mL) |
BL | 50 µL | Assay Buffer |
TS | 50 µL | Test Sample |
Prepare DNA Standards (DS), blank controls (BL), and test samples (TS) according to the layout provided in Tables 1 and 2. For a 384-well plate, use 25 μL of reagent per well instead of 50 μL.
Add 50 μL of 2X working solution to each well containing DNA Standard, blank control, and test samples to achieve a total DNA assay volume of 100 μL/well. For a 384-well plate, add 25 μL of 2X working solution to each well, resulting in a total volume of 50 μL/well.
Incubate the reaction at room temperature for 2 minutes, protected from light.
Monitor the fluorescence intensity with a fluorescence plate reader at Ex/Em = 490/530 nm (cut off at 515nm).
For data analysis, we recommend using the Online Linear Regression Calculator available here. This tool facilitates linear, logarithmic, and semi-log regression analysis to help interpret your experimental results.
Safety data sheet