Microplate fluorescence assay for the quantification of double stranded DNA using SYBR Green I dye

被引：56
|
作者：
Leggate, Johanna [1 ]
Allain, Ray [1 ]
Isaac, Leah [1 ]
Blais, Burton W. [1 ]
机构：
[1] Canadian Food Inspect Agcy, Ottawa Lab Carling, CEF, Ottawa, ON K1A 0C6, Canada
来源：
BIOTECHNOLOGY LETTERS | 2006年 / 28卷 / 19期
关键词：
DNA quantification; microplate fluorescence assay; SYBR Green I;
D O I：
10.1007/s10529-006-9128-1
中图分类号：
Q81 [生物工程学（生物技术）]; Q93 [微生物学];
学科分类号：
071005 ; 0836 ; 090102 ; 100705 ;
摘要：
A high-throughput, 96-well microplate fluorescence assay (MFA) was developed for DNA quantification using the double-stranded DNA-binding dye SYBR Green I. Samples mixed with SYBR Green I in the wells of a microtiter plate produced fluorescence in proportion with DNA concentration which was measured using a fluorescence plate reader. The performance characteristics of the assay were compared with spectrophotometric quantification based on ultraviolet absorption and the Hoefer DyNA Quant assay utilizing the fluorescent dye, Hoechst 33258. The MFA accurately quantified different types of DNA over a broad linear dynamic range of concentrations (0.25-2,500 pg/mu l), and was not affected by a variety of contaminants in the assay mixture.
引用
收藏
页码：1587 / 1594
页数：8
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