Helixyte™ Green Fluorimetric dsDNA Quantitation Kit *Optimized for Microplate Readers*


Tests: 200
Price:
$484

Overview 

Emission (nm): 525     

Excitation (nm): 490

Helixyte™ 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).

Platform

Fluorescence Microplate Reader

Excitation (nm): 490

Emission (nm): 525 

Cutoff (nm): 515  

Recommended Plate: Solid Black

 

Components

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)

 

Example protocol

At a glance

Protocol summary

  1. Add 100 µL dsDNA standards or test samples
  2. Add 100 µL Helixyte Green™ working solution
  3. Incubate at RT for 5-10 minutes
  4. 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

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.

References

 

Inhibitors of Streptococcus pneumoniae surface endonuclease EndA discovered by high-throughput screening using a PicoGreen fluorescence assay
Authors: Peterson EJ, Kireev D, Moon AF, Midon M, Janzen WP, Pingoud A, Pedersen LC, Singleton SF.
Journal: J Biomol Screen (2013): 247

Validation of a PicoGreen-based DNA quantification integrated in an RNA extraction method for two-dimensional and three-dimensional cell cultures
Authors: Chen Y, Sonnaert M, Roberts SJ, Luyten FP, Schrooten J.
Journal: Tissue Eng Part C Methods (2012): 444

Characterization of PicoGreen interaction with dsDNA and the origin of its fluorescence enhancement upon binding
Authors: Dragan AI, Casas-Finet JR, Bishop ES, Strouse RJ, Schenerman MA, Geddes CD.
Journal: Biophys J (2010): 3010

Comparison of SYBR Green I-, PicoGreen-, and [3H]-hypoxanthine-based assays for in vitro antimalarial screening of plants from Nigerian ethnomedicine
Authors: Abiodun OO, Gbotosho GO, Ajaiyeoba EO, Happi CT, Hofer S, Wittlin S, Sowunmi A, Brun R, Oduola AM.
Journal: Parasitol Res (2010): 933

Metal-enhanced PicoGreen fluorescence: application to fast and ultra-sensitive pg/ml DNA quantitation
Authors: Dragan AI, Bishop ES, Casas-Finet JR, Strouse RJ, Schenerman MA, Geddes CD.
Journal: J Immunol Methods (2010): 95

Quantification of dsDNA using the Hitachi F-7000 Fluorescence Spectrophotometer and PicoGreen dye
Authors: Moreno LA, Cox KL.
Journal: J Vis Exp. (2010)

Development and characterization of a novel host cell DNA assay using ultra-sensitive fluorescent nucleic acid stain "PicoGreen"
Authors: Ikeda Y, Iwakiri S, Yoshimori T.
Journal: J Pharm Biomed Anal (2009): 997

Enhanced DNA dynamics due to cationic reagents, topological states of dsDNA and high mobility group box 1 as probed by PicoGreen
Authors: Noothi SK, Kombrabail M, Kundu TK, Krishnamoorthy G, Rao BJ.
Journal: FEBS J (2009): 541

Factors affecting quantification of total DNA by UV spectroscopy and PicoGreen fluorescence
Authors: Holden MJ, Haynes RJ, Rabb SA, Satija N, Yang K, Blasic JR, Jr.
Journal: J Agric Food Chem (2009): 7221

Label-free DNA sequence detection with enhanced sensitivity and selectivity using cationic conjugated polymers and PicoGreen
Authors: Ren X, Xu QH.
Journal: Langmuir (2009): 43

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