Single-stranded sequencing using microfluidic reactors (SISSOR)

描述（由申请人提供）：在快速和低成本获取基因组信息方面的最新进展已经改变了生物医学和环境研究的许多方面。临床测序应用已经出现，并开始触及我们的日常生活。虽然以前认为无法实现的1000美元基因组的目标在未来2-3年内已经成为一个难以实现的目标，但在准确性和长期连续性方面仍然存在重大挑战，这两者对基因组测序的临床适用性以及许多非医学应用都有直接影响。在这个项目中，我们将开发SISSOR（单链测序使用微流控反应器），目标是以10-9或更低的误差率测序哺乳动物大小的基因组，单倍型N50至少为10 Mb，花费1000美元。此外，SISSOR可以从单个细胞开始，提供在异质组织（如癌症和大脑）中解剖体细胞突变的能力，并从难以培养的生物体中重新提取基因组信息。我们没有提出一种全新的测序方法，而是选择开发一种专注于前端制备的集成设备，与现有的合成测序化学相结合，为在4年的开发周期内实现上述目标提供了高度现实的途径。

DESCRIPTION (provided by applicant): Recent advances in acquiring genome information quickly and inexpensively have transformed many aspects of biomedical and environmental research. Clinical sequencing applications have emerged, and are at the beginning of touching our daily life. While the previously thought unreachable goal of $1,000 genome has become a difficult-to-miss target within the next 2-3 years, major challenges still remain in terms of both accuracy and long-range continuity, both have direct implications in the clinical applicability of genome sequencing as well as many non- medical applications. In this project we will develop SISSOR (SIngle-Stranded Sequencing using micrOfluidic Reactors), with the goal of sequencing a mammalian-size genome at the consensus error rate of 10-9 or lower, with a haplotype contig N50 of at least 10 Mb for $1,000. In addition, SISSOR can start with one single cell, providing the capability of dissecting somatic mutations in heterogeneous tissues (such as cancers and brain), and extracting genome information de novo from difficult-to-culture organisms. Instead of proposing a completely new sequencing method, we chose to develop an integrative device focusing on the front-end preparation, which, in combination with the existing sequencing-by-synthesis chemistry, provides a highly realistic path to achieve the above goal within a 4-year development cycle.