Enzyme-mediated single-nucleotide variation detection at room temperature with high discrimination factor

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Abstract

We demonstrate a new powerful tool to detect single-nucleotide variation in DNA at room temperature with high selectivity, based on predetermined specific interactions between Lambda exonuclease and a chemically modified DNA substrate structure which comprises two purposefully introduced mismatches and a covalently attached fluorophore. The fluorophore not only acts as a signal reporter in the detection system, but also plays a notable role in the specific molecular recognition between the enzyme and the probe/target hybrid substrate. The method is single-step, rapid, and can be easily adapted to different high-throughput micro-devices without the need for temperature control.

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Wu, T., Xiao, X., Zhang, Z., & Zhao, M. (2015). Enzyme-mediated single-nucleotide variation detection at room temperature with high discrimination factor. Chemical Science, 6(2), 1206–1211. https://doi.org/10.1039/c4sc03375b

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