公开(公告)号：US11859171B2

公开(公告)日：2024-01-02

申请号：US14784605

申请日：2014-04-17

Applicant: Agency for Science, Technology and Research

Inventor： Stephen R. Quake ,  William F. Burkholder ,  Lewis Z. Hong

IPC: C12Q1/68 ,  C12N15/10 ,  C12Q1/6869

CPC classification number: C12N15/1065 ,  C12Q1/6869 ,  C12Q1/6869 ,  C12Q2531/125 ,  C12Q2535/122 ,  C12Q2537/143 ,  C12Q2537/165 ,  C12Q2563/185

Abstract: The present invention provides an approach to increase the effective read length of commercially available sequencing platforms to several kilobases and be broadly applied to obtain long sequence reads from mixed template populations. A method for generating extended sequence reads of long DNA molecules in a sample, comprising the steps of: assigning a specific barcode sequence to each template DNA molecule in a sample to obtain barcode-tagged molecules; amplifying the barcode-tagged molecules to obtain barcode-containing fragments; juxtaposing the barcode-containing fragments to random short segments of the original DNA template molecule during the process of generating a sequencing library to obtain demultiplexed reads; and assembling the demultiplexed reads to obtain extended sequence reads for each DNA template molecule, is disclosed.

公开(公告)号：US20160076023A1

公开(公告)日：2016-03-17

申请号：US14784605

申请日：2014-04-17

Applicant: AGENCY FOR SCIENCE, TECHNOLOGY AND RESEARCH

Inventor： Stephen R. Quake ,  William F. Burkholder ,  Lewis Z. Hong

IPC: C12N15/10

CPC classification number: C12N15/1065 ,  C12Q1/6869 ,  C12Q2531/125 ,  C12Q2535/122 ,  C12Q2537/143 ,  C12Q2537/165 ,  C12Q2563/185

Abstract: The present invention provides an approach to increase the effective read length of commercially available sequencing platforms to several kilobases and be broadly applied to obtain long sequence reads from mixed template populations. A method for generating extended sequence reads of long DNA molecules in a sample, comprising the steps of: assigning a specific barcode sequence to each template DNA molecule in a sample to obtain barcode-tagged molecules; amplifying the barcode-tagged molecules; fragmenting the amplified barcode-tagged molecules to obtain barcode-containing fragments; juxtaposing the barcode-containing fragments to random short segments of the original DNA template molecule during the process of generating a sequencing library to obtain demultiplexed reads; and assembling the demultiplexed reads to obtain extended sequence reads for each DNA template molecule, is disclosed. Also disclosed are methods systems and software for assembling paired end sequence reads to produce extended reads.

Abstract translation: 本发明提供了将商业上可获得的测序平台的有效阅读长度增加到几千碱基的方法，并广泛应用于从混合模板种群获得长序列读数。 一种用于产生样品中长DNA分子的延伸序列读取的方法，包括以下步骤：将特定条形码序列分配给样品中的每个模板DNA分子以获得条形码标记的分子; 扩大条形码标签分子; 将扩增的条形码标签分子片段化，以获得含条形码的片段; 在生成测序文库的过程中将含有条形码的片段与原始DNA模板分子的随机短片段并置，以获得解复用的读数; 并且组合解复用的读取以获得每个DNA模板分子的扩展序列读取。 还公开了用于组装配对末端序列读取以产生扩展读取的方法系统和软件。