DNA labeling for improved synthetic long reads

DNA 标记可改善合成长读长

• 批准号: 10081920
• 负责人: Jonathan Alden
• 金额: $ 32.99万
• 依托单位: ESPER BIOSCIENCES, INC.
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-22 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10081920
• 关键词: Address; Area; Bar Codes; Communities; Computational algorithm; Computer software; Custom; DNA; DNA Sequence; DNA sequencing; Data; Devices; Escherichia coli; Future; Genetic; Genome; Grant; Growth; Haplotypes; Instruction; Introns; Label; Length; Libraries; Link; Messenger RNA; Methods; Modeling; Molecular; Molecular Biology; Phase; Plasmids; Play; Positioning Attribute; Procedures; Protein Isoforms; RNA Splicing; Research Personnel; Role; Route; Sampling; Scientist; Site; Spectrum Analysis; System; Technology; Transcript; Transposase; Variant; Vial device; Work; Writing; contig; cost; design; gel electrophoresis; improved; instrumentation; nanopore; next generation; next generation sequencing; single cell sequencing; single molecule; single molecule real time sequencing; synthetic genomics; tool; transcriptomics

Project Summary Long-read sequencing has the potential to greatly simplify sequencing, helping to accelerate scientists’ ability to perform de novo sequencing, haplotype phasing and transcriptomics. This project aims to develop a method to label DNA prior to next-generation sequencing, that maintains information about the proximity of fragments in the original strand, aiding in the downstream assembly of the sequencing data. To label the DNA, transposase will be loaded with specially-designed transposons containing barcode labels, and used to fragment the DNA, prior to next-generation sequencing. The first aim of this project is to construct the transposomes used to label the DNA. The second aim is to use these transposomes to tagment and sequence a model DNA system to demonstrate read lengths of ~50 kb using next-generation sequencing on less than a picogram of DNA. If successful, the sequencing approach developed in this grant will simplify synthetic long read sequencing, making high-accuracy and inexpensive long read sequencing more accessible to the genetics community.

项目摘要 长读序测序有可能大大简化测序，有助于加速科学家的研究。 进行从头测序、单体型定相和转录组学的能力。该项目旨在开发一个 一种在下一代测序之前标记DNA的方法，该方法保留了关于 在原始链中的片段，有助于测序数据的下游组装。为了标记DNA， 转座酶将装载含有条形码标签的特殊设计的转座子，并用于片段化 在下一代测序之前该项目的第一个目标是构建用于 来标记DNA第二个目标是使用这些转座体来标记和测序一个模型DNA系统 使用下一代测序技术，在不到1皮克的DNA上展示约50 kb的读段长度。如果 成功的，测序方法开发的这项赠款将简化合成长读序测序， 高准确性和廉价的长读段测序更容易为遗传学界所接受。