13 March 2012 Development of a simple and rapid assay for methylase activity based on DNA hairpin probe and Sybr Green I
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Abstract
Methylase is vital for a large number of biological reactions. Here we developed a new method for DNA methylase activity analysis. In this paper, a DNA hairpin probe with a sequence of 5'-GATC-3' in the stem region was designed. The 5'-GATC-3' sequence was targeted by Dam MTase and was methylated. Subsequently, restriction enzyme Dpnl recognized the site and cut it. Then the haipin probe was transformed into three single stranded DNA. This enzymatic process can be monitored by the change of SYBR green I fluorescence. The current label free assay is an useful tool for DNA methylase activity analysis due to its simplicity, speedability, and low cost.
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Yi Long, Yi Long, Xiaoming Zhou, Xiaoming Zhou, } "Development of a simple and rapid assay for methylase activity based on DNA hairpin probe and Sybr Green I", Proc. SPIE 8329, Tenth International Conference on Photonics and Imaging in Biology and Medicine (PIBM 2011), 83290C (13 March 2012); doi: 10.1117/12.918829; https://doi.org/10.1117/12.918829
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