Department of Chemistry, University of Oxford, Oxford, OX1 3TA, United Kingdom.
Nano Lett. 2009 Nov;9(11):3831-6. doi: 10.1021/nl9020232.
Nanopores are under development for the detection of a variety of analytes and the investigation of chemical reactions at the single molecule level. In particular, the analysis of nucleic acid molecules is under intense investigation, including the development of systems for rapid, low-cost DNA sequencing. Here, we show that DNA can be translocated through an engineered alphaHL protein pore at pH 11.7, a value at which dsDNA is denatured. Therefore, the alphaHL pore is sufficiently stable to entertain the possibility of direct nanopore sequencing of genomic dsDNA samples, which are more readily obtained and handled than ssDNA.
纳米孔技术正在被开发用于检测各种分析物，并在单分子水平上研究化学反应。特别是，核酸分子的分析受到了广泛关注，包括开发快速、低成本的 DNA 测序系统。在这里，我们证明 DNA 可以在 pH 值为 11.7 的工程化αHL 蛋白孔中穿过，此时双链 DNA 变性。因此，αHL 孔足够稳定，可以直接对基因组双链 DNA 样本进行纳米孔测序，与单链 DNA 相比，双链 DNA 更容易获得和处理。
