Helixyte™ Green Fluorimetric dsDNA Quantitation Kit *Optimized for Microplate Readers*
Price | |
Catalog Number | |
Unit Size | |
Quantity |
Telephone | 1-800-990-8053 |
Fax | 1-800-609-2943 |
sales@aatbio.com | |
Quotation | Request |
International | See distributors |
Shipping | Standard overnight for United States, inquire for international |
Excitation (nm) | 502 |
Emission (nm) | 522 |
H-phrase | H303, H313, H340 |
Hazard symbol | T |
Intended use | Research Use Only (RUO) |
R-phrase | R20, R21, R68 |
UNSPSC | 41116134 |
Helixyte™ Green Fluorimetric dsDNA Quantitation Kit *High Sensitivity* |
Helixyte™ Green Fluorimetric dsDNA Quantitation Kit *Optimized for Broad Dynamic Range* |
Overview | ![]() ![]() |
Excitation (nm) 502 | Emission (nm) 522 |
Platform
Fluorescence microplate reader
Excitation | 490 nm |
Emission | 525 nm |
Cutoff | 515 nm |
Recommended plate | Solid black |
Components
Example protocol
AT A GLANCE
Protocol summary
- 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
Important notes
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
For convenience, use the Serial Dilution Planner: https://www.aatbio.com/tools/serial-dilution/17650
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.
SAMPLE EXPERIMENTAL PROTOCOL
Table 1. Layout of dsDNA standards and test samples in a solid black 96-well microplate. DS= dsDNA Standards (DS1 - DS7, 2.3 to 1667 ng/mL); BL=Blank Control; TS=Test Samples.
BL | BL | TS | TS |
DS1 | DS1 | ... | ... |
DS2 | DS2 | ... | ... |
DS3 | DS3 | ||
DS4 | DS4 | ||
DS5 | DS5 | ||
DS6 | DS6 | ||
DS7 | DS7 |
Table 2. Reagent composition for each well.
Well | Volume | Reagent |
DS1 - DS7 | 100 µL | Serial Dilutions (2.3 to 1667 ng/mL) |
BL | 100 µL | TE |
TS | 100 µL | test sample |
- Prepare dsDNA 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 100 µL.
- Add 100 µL of Helixyte Green™ working solution to each well of dsDNA standard, blank control, and test samples to make the total dsDNA assay volume of 200 µL/well. For a 384-well plate, add 25 µL of BLANK assay mixture into each well instead, for a total volume of 50 µL/well.
- Incubate the reaction at room temperature for 5 to 10 minutes, protected from light.
- Monitor the fluorescence increase with a fluorescence microplate reader at Ex/Em = 490/525 nm (cut off at 515 nm).
Product Family
Name | Excitation (nm) | Emission (nm) |
Helixyte™ Green Fluorimetric ssDNA Quantitation Kit *Optimized for Microplate Readers* | 498 | 519 |
Images
References
Authors: Peterson EJ, Kireev D, Moon AF, Midon M, Janzen WP, Pingoud A, Pedersen LC, Singleton SF.
Journal: J Biomol Screen (2013): 247
Authors: Chen Y, Sonnaert M, Roberts SJ, Luyten FP, Schrooten J.
Journal: Tissue Eng Part C Methods (2012): 444
Authors: Dragan AI, Casas-Finet JR, Bishop ES, Strouse RJ, Schenerman MA, Geddes CD.
Journal: Biophys J (2010): 3010
Authors: Abiodun OO, Gbotosho GO, Ajaiyeoba EO, Happi CT, Hofer S, Wittlin S, Sowunmi A, Brun R, Oduola AM.
Journal: Parasitol Res (2010): 933
Authors: Dragan AI, Bishop ES, Casas-Finet JR, Strouse RJ, Schenerman MA, Geddes CD.
Journal: J Immunol Methods (2010): 95
Authors: Moreno LA, Cox KL.
Journal: J Vis Exp. (2010)
Authors: Holden MJ, Haynes RJ, Rabb SA, Satija N, Yang K, Blasic JR, Jr.
Journal: J Agric Food Chem (2009): 7221
Authors: Ikeda Y, Iwakiri S, Yoshimori T.
Journal: J Pharm Biomed Anal (2009): 997
Authors: Noothi SK, Kombrabail M, Kundu TK, Krishnamoorthy G, Rao BJ.
Journal: FEBS J (2009): 541
Authors: Ren X, Xu QH.
Journal: Langmuir (2009): 43