Cells-to-sequence sample preparation for next-generation sequencing

 描述（由申请人提供）：用于下一代测序（NGS）的样品制备是一个低效、复杂、多步骤的过程，难以实现自动化。测序文库的构建包括大约十几个生化操作。目前，这些步骤的自动化需要高度定制的自动化设备和训练有素的技术人员来管理工作流程。只有几种自动化产品可以解决这个问题，但这些产品只执行整个文库构建过程的一部分-它们不执行从生物样品中纯化DNA。随着NGS技术开始进入诊断领域，迫切需要一种可大大简化NGS样品制备过程的自动化系统。此外，开拓性的NGS公司和研究人员正在将测序领域推向非常长的单分子测序技术。大多数流行的DNA提取方法不产生长的基因组DNA分子，许多“第三代”测序公司正在积极寻求新的，可靠的方法来提取大的，完整的DNA分子，并将其转化为测序文库。在为NGS开发新的DNA大小选择技术的过程中，我们设想了一种用于自动化、快速纯化非常长的基因组DNA的强大技术。此外，该技术可以很容易地进行调整，也可以为传统的短读段和第三代长读段测序平台产生NGS文库。通过结合这两种能力，我们的第二阶段申请旨在生产一个系统，该系统能够在一个完全自动化的工作流程中从粗制血液样品中生产成品NGS文库。这样的系统将大大降低NGS样品制备的成本和复杂性，并加快NGS在诊断中的应用。

 DESCRIPTION (provided by applicant): Sample preparation for next-generation sequencing (NGS) is an inefficient, complex, multistep process that is difficult to automate. Sequencing library construction consists of around a dozen biochemical operations. Currently, automation of these steps requires highly customized automation equipment and highly trained technicians to manage the workflow. There are only a couple of automation products that address this problem, but these only perform part of the overall library construction process-they do not perform DNA purification from the biological sample. As NGS technology begins to move into the diagnostic world, there is an urgent need for walkaway, closed automated systems that dramatically simplify the NGS sample prep process. In addition, pioneering NGS companies and researchers are pushing the sequencing field toward very long-read, single-molecule sequencing technologies. This move to longer-read sequencing methods poses new challenges for NGS sample prep. Most popular DNA extraction methods do not produce long genomic DNA molecules, and many "third-gen" sequencing companies are actively seeking new, reliable methods for extracting large, intact DNA molecules, and converting them into sequencing libraries. In the course of developing new DNA size-selection technologies for NGS, we have conceived of a powerful technology for automated, rapid, purification of very long genomic DNA. In addition, the technology can be easily adapted to also produce NGS libraries for both traditional short-read and third-gen long-read sequencing platforms. By combining these two capabilities, our phase II application seeks to produce a system capable of producing finished NGS libraries from crude blood samples, in a single completely automated workflow. Such a system will dramatically reduce the cost and complexity of NGS sample prep, and speed the application of NGS to diagnostics.