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1. 发明授权

EP3218513B1 POLYNUCLEOTIDE AMPLIFICATION USING CRISPR-CAS SYSTEMS 有权
公开(公告)号：EP3218513B1
公开(公告)日：2018-10-31
申请号：EP15797569.9
申请日：2015-11-10
申请人： Illumina, Inc.
发明人： MANDELL, Jeffrey G.
IPC分类号： C12Q1/68 , C12N15/10 , C12N15/63 , C12N15/90
CPC分类号： C12Q1/6853 , C12N15/10 , C12N15/63 , C12N15/90 , C12Q2521/101 , C12Q2522/101
摘要： A method for amplifying a target nucleic acid including providing a system having a crRNA or a derivative thereof, and a Cas protein or a variant thereof. The crRNA or the derivative thereof contains a target-specific nucleotide region substantially complementary to a region of the target nucleic acid, and contacting the target nucleic acid with the system to form a complex.

2. 发明授权

EP3036343B1 HELICASE DEPENDENT AMPLIFICATION OF DNA MOLECULES USING NUCLEOTIDE ANALOGS 失效
公开(公告)号：EP3036343B1
公开(公告)日：2018-09-19
申请号：EP14837579.3
申请日：2014-08-14
申请人： Benner, Steven A. , Yang, Zunyi
发明人： Benner, Steven A. , Yang, Zunyi
IPC分类号： C12Q1/6853 , C12Q1/6844 , C12P19/34
CPC分类号： C12P19/34 , C12Q1/6844 , C12Q1/6853 , C12Q2521/513 , C12Q2525/101 , C12Q2525/117 , C12Q2521/101
摘要： This invention covers processes for the isothermal amplification of DNA molecules having a preselected sequence. It is based on the unexpected discovery that primers having, at some positions, adenine substituted by 2-aminopurine or diaminopurine, guanine by inosine, thymine by 2-thiothymine, and cytosine by N4-ethylcytosine (“SAMRS nucleotides”) were accepted by enzymes used in the standard helicase-dependent amplification (HDA). Further unexpected was the discovery that target nucleotides are efficiently amplified in an HDA-like process (hereinafter abbreviated as simply HDA) using substituted primers. Also discovered was the diminution of spurious products through the use of SAMRS-substituted primers.

3. 发明授权

EP3192900B1 METHOD FOR CONSTRUCTING NUCLEIC ACID SINGLE-STRANDED CYCLIC LIBRARY AND REAGENTS THEREOF 有权转让
公开(公告)号：EP3192900B1
公开(公告)日：2018-09-05
申请号：EP14901597.6
申请日：2014-10-14
申请人： MGI Tech Co., Ltd.
发明人： GENG, Chunyu , GUO, Rongrong , CHEN, Ruoying , HE, Lingyu , ZHANG, Wenwei , JIANG, Hui
IPC分类号： C12N15/10 , C12N15/66 , C12Q1/68 , C40B40/08 , C40B50/06
CPC分类号： C12Q1/6853 , B01J19/0046 , B01J2219/00529 , C12N9/22 , C12N9/93 , C12N11/06 , C12N15/10 , C12N15/1065 , C12N15/1082 , C12N15/1093 , C12N15/66 , C12Q1/6806 , C12Q1/6811 , C12Q1/6837 , C12Q1/6855 , C12Q1/686 , C12Q1/6874 , C12Q2525/191 , C12Q2565/537 , C40B40/06 , C40B40/08 , C40B50/06 , C40B50/14 , C40B60/14 , C12Q2521/525 , C12Q2535/122 , C12N15/1089 , C12Q2525/186 , C12Q2521/101
摘要： Provided in the present invention are a method for constructing a nucleic acid single-stranded cyclic library and reagents thereof. The method comprises the steps of using a transposase embedding complex to randomly break nucleic acids and connect a first linker; connecting a second linker at a gap; performing a first PCR reaction, wherein the 5' end of one of the primers has a first affinity tag, resulting in a product with two ends connected to different linker sequences; binding the product to a solid vector having a second affinity tag; degenerating and separating single strands having no affinity tag; and cyclizing the single strands.

4. 发明授权

EP2982762B1 NUCLEIC ACID AMPLIFICATION METHOD USING ALLELE-SPECIFIC REACTIVE PRIMER 有权
公开(公告)号：EP2982762B1
公开(公告)日：2018-08-01
申请号：EP13881318.3
申请日：2013-04-16
申请人： Genomictree, Inc.
发明人： AN, Sung Whan , OH, Tae Jeong
IPC分类号： C12Q1/6858
CPC分类号： C12Q1/6858 , C12Q2521/101 , C12Q2523/125
摘要： The present invention relates to a nucleic acid amplification method using an allele-specific reactive primer (ASRP) which is designed to solve problems of conventional allele-specific PCR. More specifically, the present invention relates to a detection method of a target nucleic acid, comprising a DNA polymerase having proofreading activity and a base sequence complementary to the target nucleic acid, wherein the target nucleic acid is amplified in the presence of an ASRP which is modified in such a manner that one or more bases from the base immediately in front, in the 5'-end direction, of a base that is cut off by the proofreading activity of the DNA polymerase when a non-complementary base exists at the 3' end, to the base at the 5' end, cannot serve as a primer for polymerization. The detection method using an ASRP according to the present invention is a technique with very high specificity of amplification due to the characteristics of the ASRP and the proofreading DNA polymerase, and is capable of effectively detecting mutations (point mutation, insertion, and deletion) including single nucleotide polymorphism (SNP). Furthermore, the method can also be used for detecting the presence of CpG methylation after bisulfite treatment, or for amplifying and detecting a target DNA starting with a desired base sequence from a DNA library.

5. 发明授权

EP2614156B1 CONTROL OF DNA MOVEMENT IN A NANOPORE AT ONE NUCLEOTIDE PRECISION BY A PROCESSIVE ENZYME 有权
公开(公告)号：EP2614156B1
公开(公告)日：2018-08-01
申请号：EP11823878.1
申请日：2011-09-07
申请人： The Regents of the University of California
发明人： CHERF, Gerald Maxwell , LIEBERMAN, Kathy R. , LAM, Christopher Evan , DOODY, Michael , AKESON, Mark A.
IPC分类号： C12Q1/68 , G01N33/50 , C12N15/09 , G01N33/487
CPC分类号： C12Q1/6869 , G01N33/48721 , C12Q2537/101 , C12Q2565/631 , C12Q2521/101 , C12Q2563/116
摘要： The invention herein disclosed provides for devices and methods that can detect and control an individual polymer in a mixture is acted upon by another compound, for example, an enzyme, in a nanopore. Of particular note is the stability of the system in a saline medium and to detect individual nucleotide bases in a polynucleotide in real time and which may be used to sequence DNA for many hours without change of reagents. The invention is of particular use in the fields of forensic biology, molecular biology, structural biology, cell biology, molecular switches, molecular circuits, and molecular computational devices, and the manufacture thereof.

6. 发明公开

EP3334840A1 SINGLE MOLECULE NUCLEIC ACID SEQUENCING WITH MOLECULAR SENSOR COMPLEXES 审中-公开
公开(公告)号：EP3334840A1
公开(公告)日：2018-06-20
申请号：EP16760599.7
申请日：2016-08-09
申请人： Stratos Genomics, Inc.
发明人： KOKORIS, Mark, Stamatios , MCRUER, Robert, N. , TABONE, John, C. , MACHACEK, Cara
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6869 , G01N27/44791 , G01N33/48721 , C12Q2521/101 , C12Q2521/513 , C12Q2521/543 , C12Q2563/116 , C12Q2565/133 , C12Q2565/607 , C12Q2565/631
摘要： The present disclosure relates to methods and constructs for single molecule electronic sequencing of template nucleic acids. The constructs are molecular sensor complexes which comprise a processive nucleic acid processing enzyme localized to a nanopore. Conformational changes in the enzyme induced by single nucleic acid processing events are transduced into electric signals by the nanopore, which are used to identify individual nucleotides. The methods can include the steps of providing a membrane with the nanopore and the enzyme complexed with a template nucleic acid localized proximal to an opening in the pore, contacting the enzyme with an ion conductive reaction mixture including the reagents required for nucleic acid processing, providing a voltage drop across the pore that induces ion current through the pore that is modulated by conformational changes in the enzyme, measuring current through the pore over time to detect nucleotide-dependent conformational changes in the enzyme, and identifying the type of nucleotide processed by the enzyme using current modulation characteristics, thus determining sequencing information about the nucleic acid molecule.

7. 发明公开

EP3334834A1 METHOD OF PREPARING CELL FREE NUCLEIC ACID MOLECULES BY IN SITU AMPLIFICATION 有权转让
公开(公告)号：EP3334834A1
公开(公告)日：2018-06-20
申请号：EP16835999.0
申请日：2016-08-12
申请人： Circulogene Theranostics, LLC
发明人： YEH, Chen-Hsiung
IPC分类号： C12P19/34 , C12Q1/68 , C40B40/06 , C40B50/06
CPC分类号： C12P19/34 , C12Q1/6806 , C12Q1/6844 , C12Q2521/101 , C12Q2521/131 , C12Q2521/307 , C12Q2521/319 , C12Q2521/501 , C12Q2521/531 , C12Q2525/155 , C12Q2543/101 , C12Y207/01078 , C12Y207/07007 , C12Y207/07049 , C12Y302/02027 , C12Y605/01001 , C40B40/06 , C40B50/06 , C12Q2525/179 , C12Q2525/191 , C12Q2525/207 , C12Q2527/101
摘要： Methods for in situ amplification (ISA) of cfNA, such as cfDNA, in a sample are provided wherein the cfNA in the sample is not subject to a nucleic acid purification step. The methods disclosed may be used to generate an analyzable pool of cfNA present in the sample. The analyzable pool may be used with a variety of analytical techniques to characterize the nucleic acid in the sample. Methods of diagnosis, determining a therapeutic intervention and monitoring of a subject are also provided.

8. 发明公开

EP3201355A1 MODIFIED POLYMERASES FOR IMPROVED INCORPORATION OF NUCLEOTIDE ANALOGUES 有权
标题翻译：改进的聚合物用于改进核苷类似物的引入
公开(公告)号：EP3201355A1
公开(公告)日：2017-08-09
申请号：EP15779120.3
申请日：2015-09-29
申请人： Illumina, Inc.
发明人： BOMATI, Erin , PREVITE, Michael , KELLINGER, Matthew William , CHEN, Cheng-Yao , HE, Molly
IPC分类号： C12Q1/68 , C12N9/12
CPC分类号： C12N9/1252 , C12Q1/6844 , C12Q1/686 , C12Y207/07007 , C12Q2521/101
摘要： Presented herein are polymerase enzymes for improved incorporation of nucleotide analogs, in particular nucleotides which are modified at the 3′ sugar hydroxyl, as well as methods and kits using the same.
摘要翻译：本文提供了用于改善核苷酸类似物掺入的聚合酶，特别是在3'糖羟基上修饰的核苷酸，以及使用其的方法和试剂盒。

9. 发明公开

EP3192900A1 METHOD FOR CONSTRUCTING NUCLEIC ACID SINGLE-STRANDED CYCLIC LIBRARY AND REAGENTS THEREOF 有权转让
标题翻译：构建核酸单链循环库的方法和试剂
公开(公告)号：EP3192900A1
公开(公告)日：2017-07-19
申请号：EP14901597.6
申请日：2014-10-14
申请人： BGI Shenzhen Co., Limited
发明人： GENG, Chunyu , GUO, Rongrong , CHEN, Ruoying , HE, Lingyu , ZHANG, Wenwei , JIANG, Hui
IPC分类号： C40B50/06 , C12Q1/68
CPC分类号： C12Q1/6853 , B01J19/0046 , B01J2219/00529 , C12N9/22 , C12N9/93 , C12N11/06 , C12N15/10 , C12N15/1065 , C12N15/1082 , C12N15/1093 , C12N15/66 , C12Q1/6806 , C12Q1/6811 , C12Q1/6837 , C12Q1/6855 , C12Q1/686 , C12Q1/6874 , C12Q2525/191 , C12Q2565/537 , C40B40/06 , C40B40/08 , C40B50/06 , C40B50/14 , C40B60/14 , C12Q2521/525 , C12Q2535/122 , C12N15/1089 , C12Q2525/186 , C12Q2521/101
摘要： Provided in the present invention are a method for constructing a nucleic acid single-stranded cyclic library and reagents thereof. The method comprises the steps of using a transposase embedding complex to randomly break nucleic acids and connect a first linker; connecting a second linker at a gap; performing a first PCR reaction, wherein the 5' end of one of the primers has a first affinity tag, resulting in a product with two ends connected to different linker sequences; binding the product to a solid vector having a second affinity tag; degenerating and separating single strands having no affinity tag; and cyclizing the single strands.
摘要翻译：本发明提供了构建核酸单链环状文库的方法及其试剂。该方法包括以下步骤：使用转座酶包埋复合物来随机破坏核酸并连接第一接头; 在间隙处连接第二连接体; 进行第一次PCR反应，其中一个引物的5'端具有第一亲和标签，产生两端连接于不同接头序列的产物; 将产物与具有第二亲和标签的固体载体结合; 退化并分离没有亲和标签的单链; 并环化单链。

10. 发明公开

EP3191630A4 METHOD FOR CONSTRUCTING SEQUENCING LIBRARY BASED ON BLOOD SAMPLE AND USAGE THEREOF IN DETERMINING FETAL GENETIC ABNORMALITY 有权
标题翻译： VERFAHREN ZUM AUFBAU EINER SEQUENZIERUNGSBIBLIOTHEK BASIEREND AUF EINER BLUTPROBE UND VERWENDUNG DAVON BEI DER BESTIMMUNG EINERFÖTALENGENETISCHENANOMALITÄT
公开(公告)号：EP3191630A4
公开(公告)日：2017-07-19
申请号：EP14901739
申请日：2014-09-26
申请人： BGI GENOMICS CO LTD
发明人： ZHANG YANYAN , CHEN FANG , JIANG HUI , GUO YULAI , ZENG PENG , XU XUN , YANG HUANMING , BALLINGER DENNIS G , BASHKIROV VLADIMIR I , SETHI HIMANSHU
IPC分类号： C40B50/06 , C12N15/10 , C12Q1/68 , C40B40/08
CPC分类号： C12N15/10 , B01J19/0046 , B01J2219/00529 , C12N15/1093 , C12N15/66 , C12Q1/6806 , C12Q1/6811 , C12Q1/6837 , C12Q1/6853 , C12Q1/6855 , C12Q1/686 , C12Q1/6874 , C12Q2525/191 , C12Q2565/537 , C40B40/06 , C40B40/08 , C40B50/06 , C40B60/14 , C12Q2521/525 , C12Q2535/122 , C12N15/1089 , C12Q2525/186 , C12Q2521/101
摘要： Provided in the present invention are a method for constructing a nucleic acid single-stranded cyclic library and reagents thereof. The method comprises the steps of using a transposase embedding complex to randomly break nucleic acids and connect a first linker; connecting a second linker at a gap; performing a first PCR reaction, wherein the 5' end of one of the primers has a first affinity tag, resulting in a product with two ends connected to different linker sequences; binding the product to a solid vector having a second affinity tag; degenerating and separating single strands having no affinity tag; and cyclizing the single strands.
摘要翻译：本发明提供了构建核酸单链环状文库的方法及其试剂。该方法包括以下步骤：使用转座酶包埋复合物来随机破坏核酸并连接第一接头; 在间隙处连接第二连接体; 进行第一次PCR反应，其中一个引物的5'端具有第一亲和标签，产生两端连接于不同接头序列的产物; 将产物与具有第二亲和标签的固体载体结合; 退化并分离没有亲和标签的单链; 并环化单链。

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