Branton Daniel, Deamer David W, Marziali Andre, Bayley Hagan, Benner Steven A, Butler Thomas, Di Ventra Massimiliano, Garaj Slaven, Hibbs Andrew, Huang Xiaohua, Jovanovich Stevan B, Krstic Predrag S, Lindsay Stuart, Ling Xinsheng Sean, Mastrangelo Carlos H, Meller Amit, Oliver John S, Pershin Yuriy V, Ramsey J Michael, Riehn Robert, Soni Gautam V, Tabard-Cossa Vincent, Wanunu Meni, Wiggin Matthew, Schloss Jeffery A
Department of Molecular and Cell Biology, Harvard University, Cambridge, Massachusetts 02138, USA.
Nat Biotechnol. 2008 Oct;26(10):1146-53. doi: 10.1038/nbt.1495.
A nanopore-based device provides single-molecule detection and analytical capabilities that are achieved by electrophoretically driving molecules in solution through a nano-scale pore. The nanopore provides a highly confined space within which single nucleic acid polymers can be analyzed at high throughput by one of a variety of means, and the perfect processivity that can be enforced in a narrow pore ensures that the native order of the nucleobases in a polynucleotide is reflected in the sequence of signals that is detected. Kilobase length polymers (single-stranded genomic DNA or RNA) or small molecules (e.g., nucleosides) can be identified and characterized without amplification or labeling, a unique analytical capability that makes inexpensive, rapid DNA sequencing a possibility. Further research and development to overcome current challenges to nanopore identification of each successive nucleotide in a DNA strand offers the prospect of 'third generation' instruments that will sequence a diploid mammalian genome for approximately $1,000 in approximately 24 h.
基于纳米孔的设备提供了单分子检测和分析能力,这是通过在电场作用下驱动溶液中的分子穿过纳米级孔隙来实现的。纳米孔提供了一个高度受限的空间,在这个空间内,可以通过多种方式之一对单个核酸聚合物进行高通量分析,并且在狭窄孔隙中能够实现的完美连续性确保了多核苷酸中核碱基的天然顺序反映在检测到的信号序列中。无需扩增或标记就可以识别和表征千碱基长度的聚合物(单链基因组DNA或RNA)或小分子(例如核苷),这种独特的分析能力使得廉价、快速的DNA测序成为可能。为克服当前DNA链中连续核苷酸纳米孔识别面临的挑战而开展的进一步研发工作,为“第三代”仪器带来了希望,这种仪器将能够在约24小时内以约1000美元的成本对二倍体哺乳动物基因组进行测序。