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

基本信息

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
项目类别：
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

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

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项目摘要

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中。现在大多数临床测序是使用短读针对的测序方法进行的，以检测突变基因组的蛋白质编码区域。但是，简读测序方法不太适合检测涉及大于100碱基对的DNA段的变化，也无法检测在染色体上相距几百个以上的序列多态性的排列。这种长期基因组分析变得越来越重要，新的长阅读测序已经开发了可以解决这些技术问题的技术。但是，新的长阅读测序方法比普遍使用的短阅读测序贵大约10倍方法。我们的建议旨在开发一种可以隔离特定长基因组DNA的仪器系统生物样品的片段（长度为100,000至100万碱基对），从而提供了新的有针对性的长阅读测序样品制备的经济方法。拟议的系统是针对的对于强大的样本吞吐量，大量基因组中心的自动化处理，临床实验室。