中国生物工程杂志

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.