SBIR Phase I: Rapid, End-to-end Sample Preparation for Sequencing Applications

这个小企业创新研究（SBIR）第一阶段项目的更广泛的影响/商业潜力包括改进制备用于测序的脱氧核糖核酸（DNA）的方法。在对生物样本进行测序之前，DNA必须从细胞中解放出来，与蛋白质和其他不需要的碎片分离，然后与特殊缓冲液和化学试剂混合。这项技术任务目前是由训练有素的科学家进行的，他们主要使用人工操作样本和昂贵的设备。本项目提出的替代方法将通过显著减少准备时间来加快基因组测序的诊断速度，同时通过自动化使准备工作更加可靠。值得注意的是，该方法有望在不减少DNA长度的情况下制备DNA；因此，该过程应该非常适合为新兴的长读测序技术准备样品。这些改进有可能减少与DNA测序相关的负担和成本，从而将DNA测序技术的好处扩大到更广泛的社会阶层。这个小企业创新研究（SBIR）第一阶段项目依赖于一个将基因组物质捕获在一个小型流动池中的过程，通过这个流动池同时施加电场和压力驱动的流动。该过程对DNA链或（核糖核酸）RNA具有高度选择性；进入流细胞的蛋白质和其他不需要的碎片穿过。而且，这个过程是比较温和的，所以样品的长度不应该因此而缩短。该项目将通过以下方式将该技术推向市场：(1)完成墨盒的设计细节和制造，包括材料选择的评估；(2)建立与墨盒对接的定制研发平台；(3)开发DNA样品提取和转移方法；(4)利用混合和加热，同时切换电流体捕获，制定库制备方案；(5)将整个制备工艺整合到单个药筒中并验证工艺性能。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

The broader impact/commercial potential of this Small Business Innovation Research (SBIR) Phase I project consists of advancing methods for preparing deoxyribonucleic acid (DNA) for sequencing. Prior to sequencing a biological sample, DNA must be liberated from cells and separated from proteins and other unwanted debris, and then mixed with specialty buffers and chemical agents. This skilled task is currently carried out by trained scientists using largely manual manipulations of the samples and expensive equipment. The alternative method proposed in this project will speed-up diagnoses from genomic sequencing by significantly reducing preparation time while also making preparation more reliable through automation. Notably, the proposed approach is expected to prepare DNA without reducing its length; consequently, the process should be ideally suited for preparing samples for emerging long-read sequencing technologies. These improvements have the potential to decrease the burden and costs associated with DNA sequencing, hence expanding the benefits of DNA sequencing technology to wider segments of society.This Small Business Innovation Research (SBIR) Phase I project relies upon a process for trapping genomic material in a small flow cell through which an electric field and pressure-driven flow are simultaneously applied. The process is highly selective towards strands of DNA or (ribonucleic acid) RNA; proteins and other, unwanted debris that enters the flow cell passes through. Also, the process is relatively gentle, so the length of sample should not be shortened as a result. This project will advance the technology to the marketplace by: (1) Completing cartridge design details and fabrication, including evaluation of material options; (2) Building a custom research and development platform for interfacing with the cartridges; (3) Developing methods for DNA sample extraction and transfer; (4) Developing the library preparation protocols using mixing and heating while toggling the Electro-Hydrodynamic Trapping; and, (5) Integrating the entirety of the preparation process into a single cartridge and validating the process performance.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.