Quantifying DNA concentrations using fluorometry: a comparison of fluorophores.
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.
Rengarajan
Kalpana
K
Department of Ophthalmology, Emory Eye Center, Emory University, Atlanta, GA 30322, USA.
Cristol
Stphen M
SM
Mehta
Milan
M
Nickerson
John M
JM
D000818:Animals; D052160:Benzothiazoles; D006690:Bisbenzimidazole / Q000737:chemistry*; D002417:Cattle; D004247:DNA / Q000032:analysis*; D003959:Diamines; D004996:Ethidium / Q000737:chemistry*; D005456:Fluorescent Dyes / Q000737:chemistry*; D007202:Indicators and Reagents; D009930:Organic Chemicals; D011804:Quinolines; D012680:Sensitivity and Specificity; D012996:Solutions; D013050:Spectrometry, Fluorescence / Q000379:methods
D052160:Benzothiazoles; D003959:Diamines; D005456:Fluorescent Dyes; D007202:Indicators and Reagents; D009930:Organic Chemicals; C099476:PicoGreen; D011804:Quinolines; D012996:Solutions; C098022:SYBR Green I; D004247:DNA; C427203:calf thymus DNA; D004996:Ethidium; D006690:Bisbenzimidazole