Achieving Single Nucleotide Resolution to Enable DNA Flossing Through Alpha-Hemolysin

项目摘要 实现常规DNA测序有可能提高对生物过程和 以及在医学、法医学和环境调查方面的援助。然而，潜在的 测序的未来好处需要降低成本和速度，并改进仪器设备 相对于当前商业可用技术的可获得性。在各种各样的下一代DNA中 随着测序技术的不断发展，纳米孔链测序已经成为一种非常有前途的 另类选择。链测序的优点是它是一种单分子技术，能够 长(千碱基)阅读，需要最少的样品准备和没有样品标记，使用相对 廉价的硬件和生化试剂，是高效序列覆盖的高通量，以及 利用直接读数，有可能读出表观遗传修饰和损伤以及其他序列 聚合物。因此，为了解决当前需要的改进，通过 使用纳米孔，EBS建议开发方法来实现单核苷酸的拆分，而不使用 利用ebs的专利电子技术、dna HL毛孔和新的α减速技术 机械装置。这些改进将使开发低复杂性(即不需要PCR)成为可能， 高精度(解决了错配和错配)纳米孔链测序方法。

Project Summary Enabling routine DNA sequencing has the potential to improve the understanding of biological processes and relationships as well as aid in medicine, forensics, and environmental investigations. However, the potential future benefits of sequencing require a reduction in cost and speed, and an improvement in instrumentation accessibility relative to presently commercially available technology. Among the various next generation DNA sequencing technologies being pursued, nanopore strand sequencing has emerged as a very promising alternative. Strand sequencing has the advantages of being a single-molecule technique that is capable of long (kilobases) reads, requires minimal sample preparation and no sample labeling, uses relatively inexpensive hardware and biochemical reagents, is high-throughput for efficient sequence coverage, and utilizes direct readout with the potential to read epigenetic modifications and lesions as well as sequence other polymers. Thus, to address the current improvements needed to enable routine DNA sequencing through the use of nanopores, EBS proposes to develop methods to achieve single-nucleotide resolution without the use of processive enzymes by taking advantage of EBS’ proprietary electronics, αHL pores and novel DNA slowing mechanisms. These advancements will enable the development of a low complexity (i.e. no PCR needed), high accuracy (indels and mismatches are resolved) nanopore strand sequencing approach.