Quantifying DNA concentrations using fluorometry: A comparison of fluorophores

• Type of Material: Article
• Authors: Kalpana Rengarajan, Emory University; Stephen M. Cristol, Emory University; Milan Mehta, Emory University; John Nickerson, Emory University
• Language: English
• Date: 2002-11-06
• Publisher: Molecular Vision
• Publication Version: Final Published Version
• Copyright Statement: © 2002 Molecular Vision
• License: Creative Commons Attribution-NonCommerical-NoDerivs 3.0 Unported
• Final Published Version (URL): http://www.molvis.org/molvis/v8/a49/
• Title of Journal or Parent Work: Molecular Vision
• ISSN: 1090-0535
• Volume: 2002
• Issue: 8
• Start Page: 416
• End Page: 421
• Grant/Funding Information: This work was supported by grants from the National Eye Institute (R03 EY13986, R01 EY09378, P30 EY6360), Research to Prevent Blindness, Inc., and the Foundation Fighting Blindness.
• Research Categories: Health Sciences, Opthamology

Abstract

Detection of low concentrations of DNA is important in vision research because many animal models only provide scant samples of ocular tissue. Quantitative analysis of low concentrations of double stranded DNA is now feasible using fluorometry with newer fluorophores. This technique offers a rapid way to evaluate the DNA content of samples based on the measurement of fluorescence enhancement emitted by fluorophore-bound DNA and is more sensitive than absorption spectrometry. The purpose of this study was to compare the sensitivity of several different fluorophores for measuring DNA concentrations by fluorometry. Based on our studies, we conclude that SYBR Green I and PicoGreen are substantially more sensitive for quantifying DNA concentrations than ethidium bromide and Hoechst 33258.