AccuGreen™ Broad Range dsDNA Quantitation Solution is a highly sensitive and accurate fluorescence-based assay designed for quantifying double-stranded DNA (dsDNA) over a broad concentration range.
It is optimized for use with fluorescence plate readers or handheld fluorometers, offering a superior alternative to traditional UV absorbance methods such as Nanodrop. The assay is ideal for applications requiring precise DNA quantitation, including next-generation sequencing (NGS) library preparation, qPCR, and DNA fragment analysis.
Key Features: Broad Quantitation Range: Accurately measures dsDNA from low nanogram to microgram levels. High Sensitivity & Specificity: Selectively binds dsDNA, minimizing interference from RNA and single-stranded DNA. Compatibility: Works with standard fluorescence plate readers and fluorometers. Easy & Rapid Protocol: Simple mix-and-read procedure with minimal pipetting steps. Low Sample Volume Requirement: Requires only a small amount of DNA sample. Protocol for dsDNA Quantitation Using AccuGreen™ Broad Range Solution
Materials Needed: AccuGreen™ Broad Range dsDNA Quantitation Solution dsDNA standards (provided or prepared) TE buffer or other compatible diluent 96-well black or clear-bottom fluorescence plate (for plate reader) Fluorescence plate reader or fluorometer (excitation/emission ~480/520 nm) Procedure: Prepare DNA Standards and Samples:
Thaw and mix the dsDNA standards well before use. Prepare serial dilutions of the DNA sample in TE buffer or appropriate diluent, ensuring they fall within the quantification range. Prepare Working Solution:
Dilute the AccuGreen™ reagent with the provided buffer (if necessary) according to the manufacturer’s instructions. Set Up Assay Plate:
Pipette X μL of dsDNA standards into designated wells. Pipette X μL of test DNA samples into separate wells. Add Y μL of working solution to each well. Mix gently by pipetting up and down or shaking the plate. Incubation:
Allow the reaction to proceed for Z minutes at room temperature in the dark. Fluorescence Measurement:
Read the fluorescence signal at excitation/emission wavelengths of ~480/520 nm using a fluorescence plate reader or fluorometer. Data Analysis:
Generate a standard curve by plotting fluorescence intensity against known DNA concentrations. Determine unknown sample concentrations by interpolating from the standard curve. Applications: Quantification of DNA for next-generation sequencing (NGS) library prep DNA fragment analysis PCR/qPCR sample normalization DNA quality control in molecular biology workflows
