Highly sensitive restriction enzyme assay and analysis: a review |
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Authors: | Liza Lam Ryota Iino Kazuhito V Tabata Hiroyuki Noji |
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Institution: | (1) The Institute of Scientific and Industrial Research, Osaka University, Osaka 567-0047, Japan |
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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 have been integrated,
which possess prospective capabilities of developing into highly sensitive and high-throughput restriction enzyme assays.
Finally, we take a brief look at the general trends in technological development in this field by comparing the advantages
and disadvantages of performing assays at bulk, microscale and single-molecule levels.
Figure Minaturization of Restriction Enzyme Assays and DNA Stretching |
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Keywords: | Bioassays Single-molecule Miniaturization Highly sensitive Restriction enzyme |
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