Targeted long-read sequencing sample preparation using Cas9 nucleases

项目摘要 DNA测序越来越多地用于遗传疾病和肿瘤学的临床研究。目前 大多数临床测序是使用短读靶向测序方法来进行的， 基因组的蛋白质编码区。然而，短读段测序方法并不很适合于 检测涉及长度大于100个碱基对的DNA片段的改变，也不能检测 染色体上相隔几百个碱基的序列多态性排列。 这种长距离基因组分析变得越来越重要，新的长读段测序技术 已经开发了能够解决这些技术问题的技术。然而，新的长期阅读 测序方法比常用的短读测序方法贵大约10倍 方法.我们的建议旨在开发一种仪器系统，可以分离特定的长基因组DNA 片段（长度为100，000至100万个碱基对），从而提供了一种新的 这是一种用于靶向长读序测序样品制备的经济方法。拟议的系统旨在 用于在大批量基因组中心进行稳健、高样本通量、自动化处理， 临床实验室

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.