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/商业潜力包括先进的制备脱氧核糖核酸（DNA）测序的方法。在对生物样品进行测序之前，DNA必须从细胞中释放出来，并与蛋白质和其他不需要的碎片分离，然后与专用缓冲液和化学试剂混合。这项技术性的工作目前由训练有素的科学家进行，主要使用手工操作样品和昂贵的设备。该项目提出的替代方法将通过显着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更加可靠。值得注意的是，所提出的方法预计将在不减少其长度的情况下制备DNA;因此，该方法应该非常适合为新兴的长读序测序技术制备样品。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测序技术的好处扩展到更广泛的社会阶层。这个小型企业创新研究（SBIR）第一阶段项目依赖于一种将基因组材料捕获在小型流动池中的过程，通过该流动池同时施加电场和压力驱动流动。该过程对DNA或（核糖核酸）RNA链具有高度选择性;进入流动池的蛋白质和其他不需要的碎片通过。此外，该过程相对温和，因此样品的长度不应因此缩短。该项目将通过以下方式将该技术推向市场：（1）完成盒设计细节和制造，包括评估材料选项;（2）建立与盒连接的定制研究和开发平台;（3）开发DNA样品提取和转移的方法;（4）开发在切换电-流体动力学捕获的同时使用混合和加热的文库制备方案;（5）开发与盒连接的定制研究和开发平台。以及，（5）将整个制备过程集成到单个盒中并验证过程性能。该奖项反映了NSF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而被认为值得支持。

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.