Efficient Creation of Long-Template Libraries for Next-Generation Sequencing

高效创建用于下一代测序的长模板库

基本信息

批准号：
9049170
负责人：
Michael L. Metzker
金额：
$ 14.68万
依托单位：
REDVAULT BIOSCIENCES, LP
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-01-14 至 2017-02-13
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9049170
关键词：
Address Agriculture Amaze Area Biological Sciences Chemistry Clinical Communities Complex Couples Custom DNA DNA Library DNA Sequence Data Development Diagnostic Disadvantaged Disease Enzymes Forensic Medicine Foundations Generations Genome Genomic DNA Genomic medicine Genomics Glean Health History of Medicine Human Hybridization Array Investigation Lead Length Libraries Methodology Methods Microfluidics Natural History New England Nucleic Acids Nucleotides Phase Play Population Dynamics Preparation Process Productivity Protein Array Protein Hybridization RNA RNA-Protein Interaction Reading Reagent Replicon Research Research Personnel Role Running Sampling Solid Structure Surface Systems Biology Technology Variant comparative genomics cost density design genome analysis genome-wide human DNA sequencing improved innovation innovative technologies method development next generation sequencing public health relevance rapid detection reference genome screening sequencing platform small molecule structural genomics technology development tool tumor progression tumorigenesis wasting

项目摘要

DESCRIPTION (provided by applicant): The advent of next-generation sequencing (NGS) technologies has allowed researchers to make unprecedented progress in the analysis of genomes, the results of which have profound implications in the realms of human health, agricultural productivity, and (natural history). The ability to fully characterize genomes remains unrealized however due in large part to our inability to reconcile sequence information over long distances. Short fragmented DNA libraries and their subsequent alignment to reference genomes, while highly useful in identifying single nucleotide variants (SNV), make de novo genome assembly and genomic structural analysis difficult. Long template NGS sequencing offers a potential solution but suffers from complex and expensive library preparation strategies. Such strategies frequently require significant amounts of input DNA due to complicated and inefficient manipulation steps. We have proposed an approach for creating long template libraries that addresses these issues with a simple, efficient, and relatively inexpensive method involving the innovative pairing of long dumbbell templates with solid-phase rolling circle replication. The successful development of this technology would greatly enhance the research community's ability to interrogate genomes, particularly those with large structural variation which likely plays a significant role in oncogenesis and tumor progression.

描述（由申请人提供）：下一代测序（NGS）技术的出现使研究人员在基因组分析方面取得了前所未有的进展，其结果对人类健康、农业生产力和（自然历史）仍然具有充分表征基因组的能力。然而，这在很大程度上是由于我们无法协调长距离的序列信息及其随后与参考基因组的比对，虽然在识别单核苷酸变异（SNV）、从头基因组组装和基因组结构分析方面非常有用，但尚未实现。长模板 NGS 测序提供了一种潜在的解决方案，但由于操作步骤复杂且效率低下，这种策略通常需要大量的输入 DNA，因此成本高昂。问题很简单，长哑铃模板与固相滚环复制的创新配对是一种高效且相对便宜的方法，该技术的成功开发将大大增强研究界研究基因组的能力，特别是那些可能发挥重要作用的结构变异较大的基因组。在肿瘤发生和肿瘤进展中。