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A method for quantification of double strand DNA using SYBR Green I dye |
刘歆 Liu Xin Jiang-feng GUO |
浙江理工大学生命科学学院生物工程研究所 |
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Abstract Abstract An accurate and high-throughput fluorescence based method was developed for quantification of double strand DNA (dsDNA) using SYBR Green I dsDNA-binding dye. A series of diluted genomic DNA and ?DNA were mixed with SYBR Green I (4×), and the fluorescence signal was measured with real-time PCR instrument and calibrated with ROX (1×) as reference dye. The performance characteristics of the method were compared with spectrophotometric quantification based on ultraviolet absorption. The new dsDNA quantification using SYBR Green I exhibited linearity over a range of ?DNA concentrations from 0.015 to 2 ng/?l and was about 100-times more sensitive and accurate than spectrophotometric quantification. The new dsDNA quantification allowed DNA sample may contain impurities and only required a small quantity of tissue sample. It could be widely used in molecular biological and diagnostic applications as an accurate and high-throughput assay tool.
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Received: 28 November 2007
Published: 25 January 2008
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