Development and application of droplet microfluidics for high-throughput single-cell and single-molecule genomics

高通量单细胞和单分子基因组学液滴微流控的开发和应用

• 批准号: RGPIN-2015-04343
• 负责人: Rodrigue, Sébastien
• 金额: $ 2.77万
• 依托单位: Université de Sherbrooke
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2015
• 资助国家: 加拿大
• 起止时间: 2015-01-01 至 2016-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=589737
• 关键词: Development; application; droplet; microfluidics; throughput

Genome sequencing is an extremely powerful tool for understanding biology. However, a relatively large amount of DNA, usually obtained by growing a large clonal population of cells under laboratory conditions, is required for this approach to work. This is difficult, or simply impossible, for the vast majority (>99%) of microorganisms that resist cultivation attempts. The overall objective of this proposal is to develop a novel high-throughput method for rapidly identifying and sequencing individual DNA molecules or complete bacterial genomes from targeted single-cells of interest within a large population. By taking advantage of droplet-based microfluidics, in which every sub-nanoliter droplet is equivalent to a reaction tube that can be manipulated, it will be possible to dramatically improve current single-cell whole-genome amplification and metagenomics methods while offering exciting new possibilities such as ultra high-throughput labeling and retrieving of droplets based on the presence or absence of specific genetic markers.

基因组测序是理解生物学的一个非常强大的工具。然而，这种方法需要相对大量的DNA，通常是通过在实验室条件下培养大量的克隆细胞群获得的。对于绝大多数（>99%）抵抗培养尝试的微生物来说，这是困难的，或者根本不可能的。该提案的总体目标是开发一种新的高通量方法，用于快速鉴定和测序单个DNA分子或完整的细菌基因组，这些DNA分子或完整的细菌基因组来自大群体中的目标单细胞。通过利用基于液滴的微流体技术，其中每个亚纳升液滴相当于可以操作的反应管，将有可能显着改善当前的单细胞全基因组扩增和宏基因组学方法，同时提供令人兴奋的新可能性，例如超高通量标记和基于特定遗传标记的存在或不存在的液滴检索。