Highly sensitive restriction enzyme assay and analysis: a review

Analytical and Bioanalytical Chemistry
Liza LamHiroyuki Noji

Abstract

Biological assays at the single molecule level are crucial to fundamental studies of DNA-protein mechanisms. In order to cater for high throughput applications, one area of immense research potential is single-molecule bioassays where miniaturized devices are developed to perform rapid and effective biological reactions and analyses. With the success of various emerging technologies for engineering miniaturized structures down to the nanoscale level, supported by specialized equipment for detection, many investigations in the field of life science that were once thought impossible can now be actively explored. In this review, the significance of downscaling to the single-molecule level is firstly presented in selected examples, with the focus placed on restriction enzyme assays. To determine the effectiveness of single-molecule restriction enzyme reactions, simple and direct analytical methods based on DNA stretching have often been reliably employed. DNA stretching can be realized based on a number of working principles related to the physical forces exerted on the DNA samples. We then discuss two examples of a nanochannel system and a microchamber system where single-molecule restriction enzyme digestion and DNA stretching ha...Continue Reading

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Citations

Jan 15, 2014·Biosensors & Bioelectronics·Jiachao YuSongqin Liu
Oct 30, 2010·Lab on a Chip·Shouichi SakakiharaHiroyuki Noji
Sep 11, 2012·Lab on a Chip·Soo Hyeon KimHiroyuki Noji
Oct 20, 2012·Angewandte Chemie·Natalia L KlyachkoAlexander V Kabanov

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