I-Corps: Amplification and Next-Generation Sequencing Library Preparation Using Picogram Quantities of Total RNA/DNA

I-Corps项目更广泛的影响/商业潜力是开发一种从少量生物材料中测序DNA/RNA的技术。高通量RNA/DNA测序被称为下一代测序（NGS），是医疗保健、医学研究和生命科学领域的一项关键技术。它用于新出现的传染病的诊断、活组织检查、产前诊断、器官移植、法医学、考古学以及基本的生物学和医学研究。然而，对少量可用的生物材料进行测序是具有挑战性的，而且大多数珍贵的生物材料通常是有限的。单细胞基因组学和空间基因组学，也涉及小数量样本，预计将是未来的重要技术，将推动对疾病，诊断和治疗的基本理解。拟议的技术可以扩大NGS的使用，并作为这一快速增长领域的使能技术。I-Corps项目的基础是开发利用下一代测序（NGS）从少量生物材料中生成DNA/RNA序列信息的技术。市场研究表明，约40%的实验室认为NGS的最大瓶颈在于“库制备方法”。提出的NGS库制备技术可扩大NGS的应用范围。与现有方法相比，该技术的效率提高了2-5倍，并且使用更少的模板（1-1/100）生成数量和质量数据。此外，所提出的技术可用于使用亚纳克（picogram）数量的模板对小rna进行测序，这是现有技术无法实现的。使用更小的样本量可以使NGS与新的生物材料一起使用，并且更准确、可靠和信息丰富，这可能会改善诊断和其他结果。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 I-Corps project is the development of a technology to sequence DNA/RNA from biological materials in small quantities. High-throughput RNA/DNA sequencing, called Next-Generation Sequencing (NGS), is a key technology in healthcare, medical research, and life sciences. It is used in the diagnosis of emerging infectious diseases, biopsies, pre-natal diagnostics, organ transplant, forensics, archaeology, and also basic biological and medical research. However, sequencing of the biomaterials available in small quantities is challenging and most precious biomaterials are usually limited in availability. Single cell genomics and spatial genomics, which also involve small quantity samples, are predicted to be important technologies in the future that will drive the basic understanding of diseases, diagnostics, and therapeutics. The proposed technology may broaden the use of NGS and serve as an enabling technology for this rapidly growing area.This I-Corps project is based on the development of technology to generate DNA/RNA sequence information using Next-Generation Sequencing (NGS) from small quantities of biological materials. Market research suggests that about ~40% of laboratories believe that the biggest bottleneck in NGS is in “library preparation methods.” The proposed NGS library preparation technology may increase the scope of NGS applications. The proposed technology has been shown to be 2-5 times more efficient and generates both quantity and quality data using a lower quantity of template (1-1/100th) compared with existing methods. In addition, the proposed technology may be used for sequencing of small-RNAs using sub-nanogram (picogram) quantity of template, which is not possible using existing technologies. This use of smaller sample sizes may allow NGS to be used with new biological materials and be more accurate, reliable, and informative, which may improve diagnostic and other results.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.