Targeted long-read sequencing sample preparation using Cas9 nucleases

使用 Cas9 核酸酶进行靶向长读长测序样品制备

• 批准号: 9980971
• 负责人: TRUETT C BOLES
• 金额: $ 79.69万
• 依托单位: SAGE SCIENCE, INC.
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-11-20 至 2021-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9980971
• 关键词: Address; Base Pairing; Biological; Cell Nucleus; Cells; Chromium; Chromosomes; Clinical; Clinical Research; Custom; DNA; DNA sequencing; Detection; Development; Digestion; Gel; Genetic Diseases; Genetic Polymorphism; Genetic Variation; Genome; Genomic DNA; Genomics; Hereditary Disease; Hour; Human; Immobilization; In Situ; Laboratories; Length; Liquid substance; Malignant Neoplasms; Measures; Methods; Mutation; Oncology; Open Reading Frames; Output; Phase; Preparation; Process; Property; Recovery; Research; Running; Sampling; Sepharose; Small Business Innovation Research Grant; System; Technology; Time; Variant; Walking; base; clinical sequencing; cost; cost effective; design; exome sequencing; gene panel; genetic testing; instrument; lymphoblastoid cell line; nuclease; operation; prototype; targeted sequencing

Project Summary DNA sequencing is increasingly being used in clinical research in genetic disease and oncology. Currently most clinical sequencing is carried out using short-read targeted sequencing methods to detect mutations in protein-coding regions of the genome. However, short-read sequencing methods are not well suited to detection of alterations involving DNA segments greater than 100 base pairs in length, nor can they detect the arrangement of sequence polymorphisms that are more than a few hundred bases apart on a chromosome. Such long-range genomic analyses are becoming increasingly important, and new long-read sequencing technologies have been developed that can address these technical problems. However, the new long-read sequencing methods are roughly 10-fold more expensive than commonly-used short-read sequencing methods. Our proposal seeks to develop an instrument system that can isolate specific long genomic DNA fragments (100,000 to 1 million base pairs in length) from biological samples, and thereby provide a new economical approach for targeted long-read sequencing sample preparation. The proposed system is intended for robust, high sample throughput, walk-away automated processing in high volume genome centers and clinical laboratories.

项目总结