Emission (nm): 525
Excitation (nm): 490Helixyte™ Green dsDNA Quantitation Assay Kit can be used for selectively detecting as little as 25 pg/ml of dsDNA in the presence of ssDNA, RNA, and free nucleotides. Helixyte™ Green exhibits large fluorescence enhancement upon binding to dsDNA. The assay is linear over three orders of magnitude and has little sequence dependence, allowing you to accurately measure DNA from many sources, including genomic DNA, viral DNA, miniprep DNA, or PCR amplification products. Helixyte™ Green dsDNA Quantitation Assay Kit is a few magnitudes more sensitive than UV absorbance readings. It is specific for dsDNA in the presence of equimolar amounts of RNA. The kit is robust with a mix and read format compatible with 96- and 384-well fluorescence-based microplate readers. It can also be used with a bench top fluorometer or a hand-held fluorescence meter (e.g., Qubit fluorometer).
Fluorescence Microplate Reader
Excitation (nm): 490
Emission (nm): 525
Cutoff (nm): 515
Recommended Plate: Solid Black
Component A: Helixyte Green™: 1 vial (1 mL, 200X in DMSO)
Component B: 20X Assay Buffer: 1 bottle (25 mL)
Component C: Calf thymus DNA Standard: 1 vial (1 mL, 100 µg/mL)
At a glance
- Add 100 µL dsDNA standards or test samples
- Add 100 µL Helixyte Green™ working solution
- Incubate at RT for 5-10 minutes
- Monitor the fluorescence at Ex/Em=490/525 nm
The following protocol is an example for quantifying dsDNA with Helixyte Green™. Allow all the components to warm to room temperature before opening. No data are available addressing the mutagenicity or toxicity of Helixyte Green™dsDNA stain. Because this reagent binds to nucleic acids, it should be treated as a potential mutagen and handled with appropriate care. The DMSO stock solution should be handled with particular caution as DMSO is known to facilitate the entry of organic molecules into tissues.
Preparation of standard solution
Add 10 µL of 100 µg/mL dsDNA stock solution (Component C) to 190 µL of Assay buffer (Component B) to have 5 µg/mL dsDNA solution, and then perform 1:3 serial dilutions to get serially diluted dsDNA standard (DS7 - DS1).
Preparation of working solution
Prepare Helixyte Green™ working solution by adding 50 μL of Helixyte Green™ (Component A) into 10 mL of Assay Buffer (Component B). Protect the working solution from light by covering it with foil or placing 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 best results, this solution should be used within a few hours of its preparation.
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