DNA sequencing using electrical conductance measurements of a DNA polymerase

Yu-Shiun Chen, Chia Hui Lee, Meng Yen Hung, Hsu An Pan, Jin-Chern Chiou, G. Steven Huang*

*此作品的通信作者

研究成果: Article同行評審

28 引文 斯高帕斯(Scopus)

摘要

The development of personalized medicine-in which medical treatment is customized to an individual on the basis of genetic information-requires techniques that can sequence DNA quickly and cheaply. Single-molecule sequencing technologies, such as nanopores, can potentially be used to sequence long strands of DNA without labels or amplification, but a viable technique has yet to be established. Here, we show that single DNA molecules can be sequenced by monitoring the electrical conductance of a phi29 DNA polymerase as it incorporates unlabelled nucleotides into a template strand of DNA. The conductance of the polymerase is measured by attaching it to a protein transistor that consists of an antibody molecule (immunoglobulin G) bound to two gold nanoparticles, which are in turn connected to source and drain electrodes. The electrical conductance of the DNA polymerase exhibits well-separated plateaux that are ∼3 pA in height. Each plateau corresponds to an individual base and is formed at a rate of ∼22 nucleotides per second. Additional spikes appear on top of the plateaux and can be used to discriminate between the four different nucleotides. We also show that the sequencing platform works with a variety of DNA polymerases and can sequence difficult templates such as homopolymers.

原文English
頁(從 - 到)452-458
頁數7
期刊Nature nanotechnology
8
發行號6
DOIs
出版狀態Published - 3 5月 2013

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