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  • 标题:Single Molecule Bioelectronics and Their Application to Amplification-Free Measurement of DNA Lengths
  • 本地全文:下载
  • 作者:O. Tolga Gül ; Kaitlin M. Pugliese ; Yongki Choi
  • 期刊名称:Biosensors
  • 电子版ISSN:2079-6374
  • 出版年度:2016
  • 卷号:6
  • 期号:3
  • 页码:29
  • DOI:10.3390/bios6030029
  • 语种:English
  • 出版社:MDPI Publishing
  • 摘要:As biosensing devices shrink smaller and smaller, they approach a scale in which single molecule electronic sensing becomes possible. Here, we review the operation of single-enzyme transistors made using single-walled carbon nanotubes. These novel hybrid devices transduce the motions and catalytic activity of a single protein into an electronic signal for real-time monitoring of the protein’s activity. Analysis of these electronic signals reveals new insights into enzyme function and proves the electronic technique to be complementary to other single-molecule methods based on fluorescence. As one example of the nanocircuit technique, we have studied the Klenow Fragment (KF) of DNA polymerase I as it catalytically processes single-stranded DNA templates. The fidelity of DNA polymerases makes them a key component in many DNA sequencing techniques, and here we demonstrate that KF nanocircuits readily resolve DNA polymerization with single-base sensitivity. Consequently, template lengths can be directly counted from electronic recordings of KF’s base-by-base activity. After measuring as few as 20 copies, the template length can be determined with <1 base pair resolution, and different template lengths can be identified and enumerated in solutions containing template mixtures.
  • 关键词:DNA polymerase; carbon nanotube sensors; single molecule enzymology; DNA sequencing DNA polymerase ; carbon nanotube sensors ; single molecule enzymology ; DNA sequencing
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