Single-stranded sequencing using microfluidic reactors (SISSOR)

描述(由申请人提供)：在快速和廉价地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。尽管之前认为1,000美元的基因组无法实现的目标已成为未来2-3年内难以错过的目标，但在准确性和长期连续性方面仍存在重大挑战，这两者都直接影响到基因组测序的临床适用性以及许多非医疗应用。在这个项目中，我们将开发SISSOR(使用微流控反应器的单链测序)，目标是以10-9或更低的普遍错误率对哺乳动物大小的基因组进行测序，单倍型重叠群N50至少为10 Mb，价格为1,000美元。此外，SISSOR可以从单个细胞开始，提供解剖异质组织(如癌症和脑)中的体细胞突变的能力，并从难以培养的生物体中从头提取基因组信息。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准备的综合设备，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一条非常现实的途径。

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.