DeNovix fluorescence assays enable dsDNA quantitation over a broad range of concentrations by employing three complimentary assay kits.
The DeNovix dsDNA Ultra High Sensitivity Assay is optimized to enable the quantification of very low concentrations of dsDNA (0.5 pg/µL to 300 pg/µL).
Like all the DeNovix dsDNA assays, the Ultra High Sensitivity Assay selectively measures concentration of dsDNA over RNA, ssDNA, or protein. The assay dye is designed for use with fluorometers and fluorescence microplate readers. The DeNovix dye has an excitation maximum at 470 nm and emission maxima at 514-567 nm. The excitation and emission spectra are shown in Figure 1.
Figure 1. Excitation and emission spectra for the DeNovix dsDNA Ultra High Sensitivity quantitation reagent in the presence of excess dsDNA.
This technical note presents typical performance data for the measurement range of the DeNovix dsDNA Ultra High Sensitivity Assay measured on DeNovix DS-11 FX fluorometers.
Materials and Methods
A dilution series of calf thymus dsDNA was prepared in TE buffer. The assay working solution was prepared by mixing 10 mL of the assay buffer with 25 µL of the dye and 100 µL of the assay enhancer solution.
For each sample, 200 µL of the working solution was added to a thin-walled, clear UV-transparent 0.5 mL PCR tube (DeNovix cat# TUBE-PCR-0.5-500). 10 μL of dsDNA was added to each tube for samples with concentrations between 0.5 pg/μL to 300 pg/μL.
Reaction solutions were mixed and incubated at room temperature for 5 minutes. Three replicates of each sample were then measured on three separate DeNovix DS-11 FX fluorometers.
The linear response of measured dsDNA and expected concentration of dsDNA measured by the Ultra High Sensitivity Assay is presented below in Figure 2.
Figure 2. Average dsDNA measured across the concentration range of the DeNovix dsDNA Ultra High Sensitivity Fluorescence Assay.
Concentration of dsDNA measured across the detection range for the DeNovix Ultra High Sensitivity Assay are shown in Table 1.
Table 1. DeNovix dsDNA Ultra High Sensitivity Assay results, measured on a DeNovix DS-11 FX.
The data presented in Table 1 and graphically represented in Figure 2 demonstrate that the DeNovix dsDNA Ultra High Sensitivity Fluorescence Assay enables measurement of dsDNA through a range of 0.5 to 300 pg/μL.
As shown in the graphic below, the DeNovix dsDNA Ultra High Sensitivity Assay provides the user with a unique ultra low concentration sample assay option.
Figure 3. Dynamic range of DeNovix dsDNA Ultra High Sensitivity Assay.
The DeNovix dsDNA Ultra High Sensitivity Assay enables accurate quantification of very low concentrations of dsDNA in an easy to use format. The simple mix and measure assay enables quantitation of dsDNA from 0.5 to 300 pg/μL. If the Ultra High Sensitivity assay does not cover the concentration range of your samples, consider using one of the alternate DeNovix dsDNA assay kits listed in Table 2.
Table 2. DeNovix dsDNA Assays.
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