公开(公告)号：WO2018073323A1

公开(公告)日：2018-04-26

申请号：PCT/EP2017/076644

申请日：2017-10-18

Applicant: OXFORD UNIVERSITY INNOVATION LIMITED

Inventor： ZHANG, Gang ,  DONNELLY, Peter ,  BOWDEN, Rory ,  HATTON, Edouard

IPC: C12Q1/68

CPC classification number: C12Q1/6844 ,  C12Q1/6853 ,  C12Q2521/307 ,  C12Q2521/327 ,  C12Q2525/179 ,  C12Q2531/119

Abstract: The present invention provides a method for the amplification of ultra-low amounts of input DNA, in particular the amount of genomic DNA which may be obtained from a single animal, plant or microbial cell, using random RNA primers and a Klenow fragment or an equivalent thereof, to obtain predominantely double stranded DNA amplification products. Further provided is a method of optimising nucleic acid sequence analysis of ultra-low amounts of DNA, said method comprising performing the DNA amplification method of the invention on an ultra-low amount of DNA prior to the nucleic acid sequence analysis of said DNA. Still further provided is a method of nucleic acid sequence analysis, said method comprising a step of sample preparation prior to the analysis step(s) in which the DNA amplification method of the invention is used to amplify an ultra-low amount of DNA in said sample.

Abstract translation: 本发明提供了使用随机RNA扩增超低量输入DNA的方法，特别是可从单个动物，植物或微生物细胞获得的基因组DNA的量的方法 引物和Klenow片段或其等同物以获得主要双链DNA扩增产物。 进一步提供了一种优化超低量DNA的核酸序列分析的方法，所述方法包括在对所述DNA进行核酸序列分析之前，在超低量的DNA上进行本发明的DNA扩增方法。 还提供了一种核酸序列分析方法，所述方法包括在分析步骤之前的样品制备步骤，其中本发明的DNA扩增方法用于扩增所述的超低量DNA 样品

公开(公告)号：WO2018033730A1

公开(公告)日：2018-02-22

申请号：PCT/GB2017/052413

申请日：2017-08-16

Applicant: TOUCHLIGHT IP LIMITED

Inventor： ADIE, Thomas ,  PORTER, Neil ,  ROTHWELL, Paul

IPC: C12Q1/68

CPC classification number: C12Q1/6844 ,  C12Q2521/113 ,  C12Q2525/151 ,  C12Q2525/301 ,  C12Q2525/307 ,  C12Q2531/119 ,  C12Q2531/125

Abstract: The present invention relates to improved processes for production of closed linear deoxyribonucleic acid (DNA), in particular cell-free enzymatic production of closed linear DNA molecules, preferably using a closed linear DNA as a template for DNA synthesis. The invention further relates to a novel closed linear DNA species, suitable for use as a template in the improved processes for production of closed linear DNA. Further, the invention pertains to the intermediate products of the processes, since this enables the production of larger quantities of closed linear DNA from the template than with methods known in the art.

Abstract translation: 本发明涉及用于产生闭合线性脱氧核糖核酸（DNA）的改进方法，特别是闭合线性DNA分子的无细胞酶促产生，优选使用闭合线性DNA作为DNA模板 合成。 本发明进一步涉及一种新的闭合线性DNA物种，其适合用作生产闭合线性DNA的改进方法中的模板。 此外，本发明涉及这些方法的中间产物，因为这使得能够比本领域已知的方法从模板产生更大量的闭合线性DNA。

公开(公告)号：WO2018013710A1

公开(公告)日：2018-01-18

申请号：PCT/US2017/041748

申请日：2017-07-12

Applicant: F. HOFFMAN-LA ROCHE AG ,  ROCHE DIAGNOSTICS GMBH ,  ROCHE SEQUENCING SOLUTIONS, INC.

Inventor： GODWIN, Brian C. ,  PLATZER, Joseph ,  MOK, Janine ,  PENKLER, Jo-Anne ,  MILLER, Bronwen

IPC: C12Q1/68

CPC classification number: C12Q1/6869 ,  C12Q1/6806 ,  C12Q1/6853 ,  C12Q2525/191 ,  C12Q2531/119 ,  C12Q2533/101 ,  C12Q2535/122 ,  C12Q2565/514

Abstract: Improved methods and compositions are provided herein for primer extension target enrichment of target polynucleotides and affinity capture. The first extension product is enriched by affinity capture using a primer or nucleotides labelled with an affinity ligant, e.g. biotin. The first extension product is displaced or degraded when a second extension primer is extended along the target strand. The second extension primer comprises a barcode region.

Abstract translation: 本文提供了用于靶多核苷酸的引物延伸靶富集和亲和捕获的改进的方法和组合物。 第一延伸产物通过使用用亲和配体标记的引物或核苷酸进行亲和捕获来富集，例如， 生物素。 当第二延伸引物沿着目标链延伸时，第一延伸产物被置换或降解。 第二个延伸引物包含条形码区域。

公开(公告)号：WO2017218864A1

公开(公告)日：2017-12-21

申请号：PCT/US2017/037819

申请日：2017-06-16

Applicant: MAYO FOUNDATION FOR MEDICAL EDUCATION AND RESEARCH

Inventor： KOCHER, Jean-Pierre A. ,  WANG, Chen

IPC: C12Q1/68 ,  G06F19/20 ,  G06F19/22

CPC classification number: C12Q1/6869 ,  C12Q1/6806 ,  C12Q1/686 ,  C12Q1/6876 ,  G16B20/00 ,  G16B30/00 ,  C12Q2531/119 ,  C12Q2535/122 ,  C12Q2537/159 ,  C12Q2563/131 ,  C12Q2537/16 ,  C12Q2537/165

Abstract: This document provides methods and materials for using low coverage whole genome sequencing techniques to assess genomes. For example, methods and materials for using targeted nucleic acid amplification and/or capture techniques in combination with low coverage whole genome sequencing techniques to obtain high coverage sequencing data for one or more pre-selected regions of a genome are provided.

Abstract translation: 本文提供了使用低覆盖度全基因组测序技术评估基因组的方法和材料。 例如，提供了使用靶向核酸扩增和/或捕获技术与低覆盖全基因组测序技术相结合以获得基因组的一个或多个预选区域的高覆盖度测序数据的方法和材料。

公开(公告)号：WO2017205304A1

公开(公告)日：2017-11-30

申请号：PCT/US2017/033885

申请日：2017-05-22

Applicant: CORNELL UNIVERSITY

Inventor： CRAIGHEAD, Harold G. ,  TIAN, Harvey C.

IPC: C12M1/00 ,  C12N15/10 ,  C12Q1/68

CPC classification number: C12N15/10 ,  B01L3/502746 ,  B01L3/502761 ,  B01L2200/0647 ,  B01L2300/0803 ,  B01L2300/0864 ,  B01L2400/0487 ,  B01L2400/086 ,  C12Q1/6844 ,  C12Q2523/303 ,  C12Q2531/119 ,  C12Q2565/629

Abstract: The present invention relates to, inter alia, a microfluidic device for performing single cell genomic DNA isolation and amplification under flow. The microfluidic device comprises a solid substrate having one or more microfluidic channel system formed therein. Each microfluidic channel system of the microfluidic device comprises: (a) an intake region comprising a single microchannel; (b) a plurality of cell segregation microchannels; (c) a cell capture site located downstream of each cell segregation microchannel; and (d) a DNA capture array positioned downstream of the cell capture site and comprising a plurality of micropillars. Also disclosed is a whole genome amplification system that includes the microfluidic device of the present disclosure, as well as a method for conducting single cell DNA analysis via on-chip whole genome amplification while under flow, and a method for multiple displacement amplification (MDA) reactions of one or more nucleic acid sequence isolated single cells.

Abstract translation: 本发明尤其涉及用于在流动下进行单细胞基因组DNA分离和扩增的微流体装置。 微流体装置包括具有在其中形成的一个或多个微流体通道系统的固体基底。 微流体装置的每个微流体通道系统包括：（a）包括单个微通道的进入区域; （b）多个细胞分离微通道; （c）位于每个细胞分离微通道下游的细胞捕获位点; 和（d）位于细胞捕获位点下游并包含多个微柱的DNA捕获阵列。 还公开了包括本公开的微流体装置的全基因组扩增系统，以及用于在流动下通过片上全基因组扩增进行单细胞DNA分析的方法，以及用于多位移扩增（MDA）的方法， 一个或多个核酸序列分离的单个细胞的反应。

公开(公告)号：WO2017176970A1

公开(公告)日：2017-10-12

申请号：PCT/US2017/026305

申请日：2017-04-06

Applicant: THE UNIVERSITY OF FLORIDA RESEARCH FOUNDATION, INC.

Inventor： WU, Chang-Yu ,  JIANG, Xiao ,  PAN, Maohua ,  LEDNICKY, John ,  THEODORE, Alexandros, Demetrios ,  FAN, Zhonghui, Hugh ,  MOHAJER, Nima, Afshar

IPC: G01N1/22 ,  C12Q1/70 ,  C12Q1/68 ,  C12Q1/02 ,  C12Q1/04 ,  C12Q1/24

CPC classification number: G01N1/22 ,  C12Q1/70 ,  G01N1/2214 ,  G01N33/54306 ,  G01N33/54366 ,  G01N33/56983 ,  G01N2001/2223 ,  G01N2001/2282 ,  C12Q2531/119 ,  C12Q2565/629 ,  C12Q2531/143

Abstract: Described herein are bioaerosol detection systems and methods of use.

Abstract translation: 本文描述了生物气溶胶检测系统和使用方法。

公开(公告)号：WO2017162765A1

公开(公告)日：2017-09-28

申请号：PCT/EP2017/056865

申请日：2017-03-22

Applicant: UNIVERSITETET I TROMSØ - NORGES ARKTISKE UNIVERSITET

Inventor： LARSEN, Atle Noralf ,  PIOTROWSKI, Yvonne ,  ASSEFA, Netsanet Gizaw ,  LANES, Olav

IPC: C12N9/12

CPC classification number: C12N9/1252 ,  C12Q1/6844 ,  C12Q2521/101 ,  C12Q2531/101 ,  C12Q2531/119 ,  C12Q2531/125 ,  C12Y207/07007

Abstract: The present invention relates to DNA polymerases. In particular, the present invention relates to DNA polymerases based on a DNA polymerase from a Psychrobacillus sp . The present invention provides an isolated DNA polymerase or an enzymatically active fragment thereof, said DNA polymerase comprising the amino acid sequence of SEQ ID NO:1 or comprising an amino acid sequence which is at least 70% identical to SEQ ID NO:1. The invention also provides nucleic acid molecules comprising a nucleotide sequence that encodes the DNA polymerase. The invention also provides a method of nucleotide polymerisation and a method of amplifying a nucleic acid in which the DNA polymerase or an enzymatically active fragment thereof is used.

Abstract translation: 本发明涉及DNA聚合酶。 特别地，本发明涉及基于来自嗜热脂肪杆菌属的DNA聚合酶的DNA聚合酶。 本发明提供了分离的DNA聚合酶或其酶促活性片段，所述DNA聚合酶包含SEQ ID NO：1的氨基酸序列或包含与SEQ ID NO：1至少70％相同的氨基酸序列。 本发明还提供了包含编码DNA聚合酶的核苷酸序列的核酸分子。 本发明还提供了一种核苷酸聚合方法和扩增其中使用了DNA聚合酶或其酶促活性片段的核酸的方法。

公开(公告)号：WO2016123692A1

公开(公告)日：2016-08-11

申请号：PCT/CA2016/000031

申请日：2016-02-04

Applicant: THE UNIVERSITY OF BRITISH COLUMBIA

Inventor： HANSEN, Carl Lars Genghis ,  ZAHN, Hans ,  HUFT, Jens ,  VAN LOENHOUT, Marinus Theodorus Johannes ,  LEUNG, Kaston ,  LIN, Bill Kengli ,  KLAUS, Anders ,  APARICIO, Samuel Alves Jana Rodrigues ,  SHAH, Sohrab Prakash

IPC: C40B50/06 ,  B81B1/00 ,  B81B7/04 ,  C12M1/34 ,  C12N15/00 ,  C12P19/34 ,  C12Q1/00 ,  C12Q1/68 ,  C40B20/00 ,  C40B30/00 ,  C40B40/06 ,  G01N33/48 ,  G01N33/50 ,  G01N33/53 ,  G06F19/10

CPC classification number: C12Q1/6869 ,  B01L3/502761 ,  B01L3/505 ,  B01L2300/0887 ,  B01L2300/123 ,  B01L2300/14 ,  C12N15/1093 ,  C12Q1/6806 ,  C12Q1/686 ,  C12Q2531/119 ,  C12Q2563/149 ,  C12Q2565/629 ,  C12Q2525/191 ,  C12Q2531/113

Abstract: Methods, devices and systems for analyzing precious samples of cells, including single cells are provided. The methods, devices, and systems in various embodiments of the invention are used to assess genomic heterogeneity, which has been recognized as a central feature of many cancers and plays a critical role in disease initiation, progression, and response to treatment. The methods devices and systems are also used to analyze embryonic biopsies for reimplantation genetic diagnosis (PGD). In one embodiment, the devices, systems and methods provided herein allow for the construction of genomic and RNA-seq libraries without a pre- amplification step.

Abstract translation: 提供了分析包括单细胞在内的珍贵细胞样品的方法，装置和系统。 本发明的各种实施方案中的方法，装置和系统用于评估基因组异质性，其被认为是许多癌症的中心特征，并且在疾病起始，进展和对治疗的反应中起关键作用。 方法设备和系统也用于分析胚胎活组织检查用于再植入遗传诊断（PGD）。 在一个实施方案中，本文提供的装置，系统和方法允许在没有预扩增步骤的情况下构建基因组和RNA-seq文库。

公开(公告)号：WO2016059473A2

公开(公告)日：2016-04-21

申请号：PCT/IB2015/002141

申请日：2015-10-13

Applicant: ABBOTT JAPAN CO., LTD ,  TOKYO INSTITUTE OF TECHNOLOGY

Inventor： KOMIYA, Ken ,  KOMORI, Makoto ,  TORU, Yoshimura

IPC: C12Q1/68

CPC classification number: C12Q1/682 ,  C12Q1/6844 ,  C12Q2525/113 ,  C12Q2525/131 ,  C12Q2525/185 ,  C12Q2531/119

Abstract: Disclosed are methods for detecting a target nucleic acid in a sample. The methods include contacting said sample, in the presence of a polymerase and an endonuclease, with a sequence conversion oligonucleotide having locked nucleic acids at select positions sufficient to decrease non-specific background signal amplification. Also disclosed are methods for detecting a target nucleic acid in a sample in which said sample is contacted, in the presence of a polymerase and an endonuclease, with a sequence conversion oligonucleotide and a signal amplifier oligonucleotide, both having locked nucleic acids at select positions sufficient to decrease non-specific background signal amplification. The disclosure also provides compositions and kits comprising such sequence conversion and signal amplifier oligonucleotides.

Abstract translation: 公开了用于检测样品中的靶核酸的方法。 所述方法包括使所述样品在聚合酶和内切核酸酶的存在下与序列转换寡核苷酸接触，所述序列转换寡核苷酸在足以降低非特异性背景信号扩增的选择位置处具有锁核酸。 还公开了在聚合酶和内切核酸酶存在下，用序列转换寡核苷酸和信号放大寡核苷酸检测其中所述样品接触的样品中的靶核酸的方法，所述序列转换寡核苷酸和信号放大寡核苷酸均在选择位置处具有锁定核酸 减少非特异性背景信号放大。 本公开还提供了包含这种序列转换和信号放大寡核苷酸的组合物和试剂盒。

公开(公告)号：WO2015171656A1

公开(公告)日：2015-11-12

申请号：PCT/US2015/029311

申请日：2015-05-05

Applicant: BAYLOR COLLEGE OF MEDICINE

Inventor： ZONG, Chenghang ,  GUNDRY, Michael ,  SHENG, Kuanwei

IPC: C12Q1/68 ,  C12P19/34

CPC classification number: C12Q1/6844 ,  C12Q1/6804 ,  C12Q1/6846 ,  C12Q2521/301 ,  C12Q2525/131 ,  C12Q2525/161 ,  C12Q2531/119

Abstract: Embodiments of the disclosure encompass methods of amplifying nucleic acid from one or more cells using MALBAC (multiple annealing and looping-based amplification cycles) primers. In particular embodiments, the nucleic acid is amplified as amplicons in a linear manner. Specific embodiments include the removal or effective destruction of nonlinearly produced amplicons.

Abstract translation: 本公开的实施方案包括使用MALBAC（多重退火和基于循环的扩增循环）引物扩增来自一个或多个细胞的核酸的方法。 在具体实施方案中，核酸以线性方式扩增为扩增子。 具体实施方案包括去除或有效破坏非线性产生的扩增子。