Integrated system for preparation of NGS libraries from crude biological samples

用于从粗生物样品制备 NGS 文库的集成系统

• 批准号: 8314280
• 负责人: TRUETT C BOLES
• 金额: $ 14.41万
• 依托单位: SAGE SCIENCE, INC.
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-07-06 至 2014-01-08
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8314280
• 关键词: Adsorption; Area; Biological; Blood; Cancer Diagnostics; Cell Nucleus; Cells; Clinical; Clinical Trials; Complex; DNA; DNA Library; DNA Sequence; Development; Devices; Diagnostic; Diagnostic tests; Exhibits; Generations; Genomics; Goals; Intervention; Leukocytes; Libraries; Manuals; Medicine; Microfluidics; Molecular Weight; Movement; Organelles; Particle Size; Phase; Preparation; Process; Property; Protocols documentation; Reaction; Research; Sampling; Small Business Innovation Research Grant; Solid; System; Technology; Whole Blood; Work; anticancer research; base; cost; cost effective; design; instrument; next generation; novel strategies; oncology; operation; particle; programs; prototype

DESCRIPTION (provided by applicant): The overall goal for the proposed SBIR program is to develop an instrument system that can accept whole blood as an input sample and produce a genomic DNA library suitable for next- generation DNA sequencing (NGS). All steps, from DNA extraction to final library construction, take place by a single automated process, without any user intervention. Because of the fully automated process, the proposed system would greatly lower the cost and labor of NGS sequencing, and accelerate movement of NGS technology into clinical diagnostic settings. Another advantage of the instrument is that it should be scalable from 100's of ml of blood down to the single cell level. Application of NGS technology to small samples is an increasingly important goal in cancer research and diagnostics. The proposed system utilizes a novel approach to microfluidic sample preparation. A single continuous-flow technology is used to manipulate and separate cells, subcellular organelles, and large genomic DNA molecules (that exhibit particle-like properties). The use of a common technology for all sample prep steps provides several benefits: 1) Multiple sequential processing steps can be accomplished in a single operation, on a single consumable device, thereby simplifying system design. 2) In multi-step processes, integration of reaction steps and post-reaction cleanup steps enables seamless, potentially zero-loss transfer of sample between processing steps. 3) Sample purification is accomplished on the basis of "particle" size alone. Differential adsorption to solid phases is not used, and sample loss due to incomplete elution is voided. 4) It is straightforward to automate the lengthy, complex sample preparation protocols that are typical of most NGS platforms, making it easier and more cost effective to move NGS technology into high throughput applications like clinical trials research and diagnostic testing. PUBLIC HEALTH RELEVANCE: The overall goal for the proposed SBIR program is to develop an instrument system that can accept whole blood as an input sample and produce a genomic DNA library suitable for next- generation DNA sequencing (NGS). All steps, from DNA extraction to final library construction, take place by a single automated process, without any user intervention. Because of the fully automated process, the proposed system would greatly lower the cost and labor of NGS sequencing, and accelerate movement of NGS technology into clinical diagnostic settings.

描述（由申请人提供）：拟议的SBIR计划的总体目标是开发一种可以接受全血作为输入样本的仪器系统，并产生适合下一代DNA测序（NGS）的基因组DNA库。从DNA提取到最终图书馆构建的所有步骤，都是通过单个自动化过程进行的，而无需任何用户干预。由于完全自动化的过程，提议的系统将大大降低NGS测序的成本和劳动，并加速NGS技术进入临床诊断环境。该仪器的另一个优点是，它应该从100毫升的血液向下扩展到单细胞水平。 NGS技术在小样本中的应用是癌症研究和诊断的越来越重要的目标。所提出的系统利用一种新的方法进行微流体样品制备。一种连续流技术用于操纵和分离细胞，亚细胞细胞器和大型基因组DNA分子（表现出类似粒子的特性）。在所有示例准备步骤中使用通用技术提供了几个好处：1）在单个可消耗设备上可以在单个操作中完成多个顺序处理步骤，从而简化了系统设计。 2）在多步骤过程中，反应步骤和反应清理步骤的整合使得在处理步骤之间的样本的无缝，可能为零损坏。 3）仅基于“粒子”大小来完成样品纯化。不使用差分吸附到固体相位，并且由于不完全洗脱而导致的样本损失变白。 4）自动化大多数NGS平台典型的冗长，复杂的样本准备协议是很简单的，这使得将NGS技术转移到诸如临床试验研究和诊断测试之类的高吞吐量应用程序中变得更加容易，更具成本效益。 公共卫生相关性：拟议的SBIR计划的总体目标是开发一种可以接受全血作为输入样本的仪器系统，并产生适用于下一代DNA测序（NGS）的基因组DNA库。从DNA提取到最终图书馆构建的所有步骤，都是通过单个自动化过程进行的，而无需任何用户干预。由于完全自动化的过程，提议的系统将大大降低NGS测序的成本和劳动，并加速NGS技术进入临床诊断环境。