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1. 发明申请

WO2019032760A1 IMPROVED METHOD TO ANALYZE NUCLEIC ACID CONTENTS FROM MULTIPLE BIOLOGICAL PARTICLES 审中-公开
公开(公告)号：WO2019032760A1
公开(公告)日：2019-02-14
申请号：PCT/US2018/045891
申请日：2018-08-09
申请人： ROOTPATH GENOMICS, INC.
发明人： PORTER, Ely , CHEN, Xi
IPC分类号： C12Q1/6806 , C12Q1/6869
CPC分类号： C12Q1/6806 , C12Q1/6869 , C12Q2521/107 , C12Q2535/122 , C12Q2563/149 , C12Q2563/159 , C12Q2563/179 , C12Q2521/325 , C12Q2522/101 , C12Q2537/161
摘要： The application provides improved methods of analyzing biological particles and their constituents, including methods of labeling at least one target nucleic acid molecule from a biological particle with a barcoded primer. The method is based on compartments (droplets) with cells and beads carrying the barcoded primers. The compartments are broken and the content contacted with polymerase, e.g. reverse transcriptase. The method is characterised in that it contains a step of inactivating the primers on the beads, which have not reacted with target, using oligos having sequences complementary to the beads; in that it contains steps for fixation reversal of fixed cells; or in that the barcoded primers are pooled after the primers have been mobilised from the beads.

2. 发明申请

WO2013160563A1 METHOD FOR COMPETITIVE ALLELE-SPECIFIC CDNA SYNTHESIS AND DIFFERENTIAL AMPLIFICATION OF THE CDNA PRODUCTS 审中-公开
标题翻译：竞争性特异性CDNA合成和CDNA产品差异放大的方法
公开(公告)号：WO2013160563A1
公开(公告)日：2013-10-31
申请号：PCT/FI2013/050470
申请日：2013-04-25
申请人： HO, Tho Huu
发明人： HO, Tho Huu
IPC分类号： C12Q1/68 , C12N15/11
CPC分类号： C12Q1/6858 , C12Q2521/107 , C12Q2525/155 , C12Q2525/161 , C12Q2531/113 , C12Q2537/161
摘要： The present invention provides a method for the detection of the presence of RNA variants comprising the step of performing a competitive c DNA synthesis comprising a first primer specific to a first RNA variant, a second primer specific to a second RNA variant, an RNA-dependent DNA polymerase, and RNA from said sample as a template, wherein said first primer and said second primer comprise an allele-specific nucleotide portion, a target-specific sequence and tag units with a common sequence and/or a discriminating sequence so that the sequence of said tag units is not complementary to said first or second RNA variant, wherein each of the c DNA products obtained from said competitive c DNA synthesis consists of the sequence of only one primer extended by the sequence complementary to one of the target RNA variants.
摘要翻译：本发明提供了检测RNA变体存在的方法，其包括进行竞争性c DNA合成的步骤，其包括对第一RNA变体特异性的第一引物，对第二RNA变体特异性的第二引物，RNA依赖性 DNA聚合酶和来自所述样品的RNA作为模板，其中所述第一引物和所述第二引物包含等位基因特异性核苷酸部分，靶特异性序列和具有共同序列和/或鉴别序列的标签单位，使得序列的所述标签单元不与所述第一或第二RNA变体互补，其中从所述竞争性cDNA合成获得的每个cDNA产物由仅与一种靶RNA变体互补的序列延伸的一个引物的序列组成。

3. 发明申请

WO2013003849A4 PROCESS AND APPARATUS FOR QUANTIFYING NUCLEIC ACID IN A SAMPLE 审中-公开
标题翻译：在样品中定量核酸的方法和装置
公开(公告)号：WO2013003849A4
公开(公告)日：2013-06-27
申请号：PCT/US2012045271
申请日：2012-07-02
申请人： BIOQUANTA SA , BIOQUANTA CORP , ASSIST PUBL HOPITAUX DE PARIS , CONTI MARC , LORIC SYLVAIN , MANIVET PHILIPPE , DELACOTTE NICOLAS , KEUMEUGNI KWEMO CARLOSSE , SALIOU BAH MAMADOU
发明人： CONTI MARC , LORIC SYLVAIN , MANIVET PHILIPPE , DELACOTTE NICOLAS , KEUMEUGNI KWEMO CARLOSSE , SALIOU BAH MAMADOU
IPC分类号： C12Q1/68 , G01N33/53
CPC分类号： C12Q1/6806 , C12Q1/68 , C12Q2537/125 , C12Q2545/113 , C12Q2563/173 , C12Q2522/101 , C12Q2537/161 , C12Q2545/114
摘要： In one aspect, the present invention concerns methods and kits for quantifying a target nucleic acid in a sample. In embodiments, a method comprises sequestrating the target nucleic acid by one or more interactors to form a sequestered conjugate that will be further labeled to form a labeled conjugate. The signal produced by the labeled conjugate is measured and correlated to the amount of the target nucleic acid. In another aspect, the present invention concerns a method for quantifying a target nucleic acid in a sample by a reaction of the nucleic acids in the sample with a nucleic acid interactor or set of interactors to form a conjugate. The conjugate is then sequestered from the rest of the sample with a molecule bound to a support, to form a sequestered conjugate. The sequestered conjugate is labeled to form a labeled conjugate. The signal produced by the labeled conjugate is measured and correlated to the amount of the target nucleic acid. In some embodiments, the target nucleic acid is cell free nucleic acids. In embodiments, the interactor binds to nucleic acids in the sample in a nonsequence specific manner.
摘要翻译：一方面，本发明涉及用于定量样品中靶核酸的方法和试剂盒。在实施方案中，方法包括通过一种或多种相互作用体螯合靶核酸以形成将被进一步标记以形成标记的缀合物的螯合缀合物。测量由标记的缀合物产生的信号并与靶核酸的量相关。在另一方面，本发明涉及通过样品中的核酸与核酸相互作用体或一组相互作用体的反应来定量样品中的靶核酸以形成缀合物的方法。然后将缀合物与其余的样品以分子结合到载体上螯合形成螯合偶联物。被隔离的缀合物被标记以形成标记的缀合物。测量由标记的缀合物产生的信号并与靶核酸的量相关。在一些实施方案中，靶核酸是无细胞核酸。在实施方案中，相互作用者以非序列特异性方式结合样品中的核酸。

4. 发明申请

WO2011000836A1 OLIGONUCLEOTIDE HYBRIDIZATION METHOD 审中-公开
标题翻译：寡核苷酸杂交方法
公开(公告)号：WO2011000836A1
公开(公告)日：2011-01-06
申请号：PCT/EP2010/059218
申请日：2010-06-29
申请人： AIT AUSTRIAN INSTITUTE OF TECHNOLOGY GMBH , KRAINER, Siegfried , FLUCH, Silvia , LEVENTE, Bodrossy , STIERSCHNEIDER, Michael
发明人： KRAINER, Siegfried , FLUCH, Silvia , LEVENTE, Bodrossy , STIERSCHNEIDER, Michael
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6832 , C12Q1/6837 , C12Q2537/161 , C12Q2527/101 , C12Q2525/107 , C12Q2527/107 , C12Q2525/121
摘要： The present invention provides a method to bind oligonucleotide molecules of a sample to an oligonucleotide probe with increased specificity comprising the steps of - providing a solid support with at least one immobilized oligonucleotide DNA probe comprising an oligonucleotide probe sequence, - in any order, contacting said probes with a sample potentially comprising oligonucleotide molecules binding said probes and contacting said probes with at least one competitive non-DNA oligonucleotide, wherein said competitive oligonucleotide comprises a sequence which is complementary to the oligonucleotide probe sequence; and/or as an alternative or in addition, in any order, contacting said probes with a sample comprising oligonucleotide molecules and contacting said oligonucleotide molecules with at least one competitive non-DNA oligonucleotide, wherein said competitive oligonucleotide comprises a sequence of the oligonucleotide probe sequence, - incubating said contacted probes with the oligonucleotide molecules of the sample and competitive oligonucleotides at a temperature of between 15°C below the melting temperature of DNA duplex oligonucleotides of the probe sequence up to said melting temperature, thereby increasing binding specificity of the oligonucleotide probe to an oligonucleotide molecule comprising a sequence complementary to said probe sequence; as well as means for performing said method.
摘要翻译：本发明提供了一种将样品的寡核苷酸分子与特异性增加的寡核苷酸探针结合的方法，包括以下步骤：提供固体支持物与至少一种包含寡核苷酸探针序列的固定寡核苷酸DNA探针， - 以任何顺序，将所述具有潜在地包含结合所述探针并使所述探针与至少一个竞争性非DNA寡核苷酸接触的寡核苷酸分子的样品的探针，其中所述竞争性寡核苷酸包含与寡核苷酸探针序列互补的序列; 和/或作为替代或另外的任何顺序，使所述探针与包含寡核苷酸分子的样品接触并使所述寡核苷酸分子与至少一个竞争性非DNA寡核苷酸接触，其中所述竞争性寡核苷酸包含寡核苷酸探针序列， - 将所述接触的探针与样品的寡核苷酸分子和竞争性寡核苷酸在比探针序列的DNA双链寡核苷酸的解链温度低15℃的温度下孵育直至所述解链温度，从而增加寡核苷酸探针的结合特异性涉及包含与所述探针序列互补的序列的寡核苷酸分子; 以及用于执行所述方法的装置。

5. 发明申请

WO2010113452A1 遺伝子型の識別方法审中-公开
标题翻译：鉴别基因组方法
公开(公告)号：WO2010113452A1
公开(公告)日：2010-10-07
申请号：PCT/JP2010/002200
申请日：2010-03-26
申请人：凸版印刷株式会社 , 山根明男 , 中山尚母 , 北野史朗
发明人：山根明男 , 中山尚母 , 北野史朗
IPC分类号： C12Q1/68 , C12N15/09 , G01N21/78 , G01N33/50 , G01N33/566
CPC分类号： C12Q1/686 , C12Q1/6858 , G01N2021/6441 , C12Q2537/161 , C12Q2535/131 , C12Q2531/119 , C12Q2525/186 , C12Q2527/125
摘要：　本願発明は、試料に含まれる遺伝子中の変異部位を含む領域を核酸増幅反応により増幅し、試料２本鎖核酸を含有する増幅反応液を得る核酸増幅工程と、前記変異部位が特定の遺伝子型であり、かつ標識物質により標識されている標準２本鎖核酸と、前記核酸増幅工程で得られた増幅反応液とを混合して競合的鎖置換反応を行い、標準２本鎖核酸と試料２本鎖核酸との間で鎖置換が生じた程度を測定することにより、前記標準２本鎖核酸と前記試料２本鎖核酸との同一性を識別する識別工程とを有し、かつ、前記競合的鎖置換反応が、ポリメラーゼによる伸長反応が抑制された条件で行われる、ＰＣＲ－ＰＨＦＡ法を利用した遺伝子型の識別方法、並びに、当該方法により遺伝子型を識別するために用いられる遺伝子型識別用キットに関する。
摘要翻译：提供了使用PCR-PHFA区分基因型的方法，所述方法包括核酸扩增过程和鉴别过程。核酸扩增方法使用核酸扩增反应来扩增样品中包含的基因的区域，所述区域包括突变位点，从而从样品获得含有双链核酸的扩增反应溶液。在鉴别过程中，将在核酸扩增过程中获得的扩增反应溶液与上述突变位点具有特定基因型的参考双链核酸混合并用标记物质标记; 然后进行竞争性链置换反应，通过测定参考双链核酸与样品双链核酸之间发生链排列的程度，确定参考双链核酸样品双链核酸是相同的。竞争性链置换反应在聚合酶延伸反应被抑制的条件下进行。还提供了用于使用该方法区分基因型的基因型鉴别试剂盒。

6. 发明申请

WO2010027484A3 ENGINEERING POLYMERASES AND REACTION CONDITIONS FOR MODIFIED INCORPORATION PROPERTIES 审中-公开
标题翻译：工程聚合物与改性掺入特性的反应条件
公开(公告)号：WO2010027484A3
公开(公告)日：2010-06-24
申请号：PCT/US2009004993
申请日：2009-09-04
申请人： PACIFIC BIOSCIENCES CALIFORNIA , PATEL PRANAV , KORLACH JONAS , BIBILLO ARKADIUSZ , BJORNSON KEITH , HANES JEREMIAH
发明人： PATEL PRANAV , KORLACH JONAS , BIBILLO ARKADIUSZ , BJORNSON KEITH , HANES JEREMIAH
IPC分类号： C12Q1/68 , C12N15/11 , C12Q1/25
CPC分类号： C12Q1/6869 , C12N9/1252 , C12Q2527/113 , C12Q2521/101 , C12Q2527/137 , C12Q2537/161 , C12Q2525/101 , C12Q2563/107
摘要： Provided are methods for enhanced sequencing of nucleic acid templates. Also provided are reaction conditions that increase branching fractions during polymerization reactions. Also provided are compositions comprising modified recombinant polymerases that exhibit branching fractions that are higher than the branching fractions of the polymerases from which they were derived. Provided are compositions comprising modified recombinant polymerases that exhibit delayed translocation relative to the polymerases from which they were derived. Also provided are compositions comprising modified recombinant polymerases that exhibit increased nucleotide or nucleotide analog residence time at an active site of the polymerase. Provided are methods for generating polymerases with the aforementioned phenotypes and methods of using such polymerases to sequence a DNA template or make a DNA. Also provided are methods and nucleic acid sequencing systems for determining which labeled nucleotide is incorporated at a site during a template-dependent polymerization reaction.
摘要翻译：提供了用于增强核酸模板测序的方法。还提供了在聚合反应期间增加支化部分的反应条件。还提供了包含修饰的重组聚合酶的组合物，其表现出比衍生它们的聚合酶的支化部分更高的支化部分。提供了包含相对于衍生它们的聚合酶显示延迟易位的修饰的重组聚合酶的组合物。还提供了包含修饰的重组聚合酶的组合物，所述修饰的重组聚合酶在聚合酶的活性位点显示增加的核苷酸或核苷酸类似物停留时间提供了产生具有上述表型的聚合酶的方法和使用这些聚合酶测序DNA模板或制备DNA的方法。还提供了用于确定哪些标记的核苷酸在模板依赖性聚合反应期间在位点处掺入的方法和核酸测序系统。

7. 发明申请

WO2008097349A2 NUCLEIC ACID ARRAY HAVING FIXED NUCLEIC ACID ANTI-PROBES AND COMPLEMENTARY FREE NUCLEIC ACID PROBES 审中-公开
标题翻译：具有固定核酸抗原探针和完全免费的核酸探针的核酸阵列
公开(公告)号：WO2008097349A2
公开(公告)日：2008-08-14
申请号：PCT/US2007/076382
申请日：2007-08-21
申请人： CNVGENES, INC. , ZAINIEV, Gafur , ZAINIEV, Inlik
发明人： ZAINIEV, Gafur , ZAINIEV, Inlik
IPC分类号： C12Q1/68 , C07H21/04
CPC分类号： C12Q1/6837 , C12Q2537/161
摘要： A process for identifying a complementary nucleic acid probe to a target nucleic acid involves forming an array of spots where each spot of the array has an immobilized nucleic acid anti-probe to which is hybridized a nucleic acid probe. The array of the anti-probe-probe complex is denatured. The nucleic acid probes are then moved into a target chamber that includes a target nucleic acid. Hybridization conditions are established to form double-stranded complexation between the target nucleic acid and nucleic acid probes in instances where the target nucleic acid has a sequence complementary. The nucleic acid probes noncomplementary to the target nucleic acid are allowed to rehybridize with anti-probes. Determining whether the anti-probe spots exposed to nucleic acid probes noncomplementary to the target nucleic acid are single stranded after exposure to noncomplementary nucleic acid probes provides information as to target nucleic acid sequence.
摘要翻译：用于鉴定靶核酸的互补核酸探针的方法包括形成阵列阵列，其中阵列的每个点具有与核酸探针杂交的固定的核酸抗探针。抗探针 - 探针复合物的阵列变性。然后将核酸探针移动到包括靶核酸的靶室中。建立杂交条件以在目标核酸具有序列互补性的情况下在靶核酸和核酸探针之间形成双链复合。允许与靶核酸不互补的核酸探针用抗探针重新杂化。确定在暴露于非互补核酸探针之后，暴露于与靶核酸不互补的核酸探针的抗探针点是单链提供关于靶核酸序列的信息。

8. 发明申请

WO2005033340A2 METHODS AND KITS FOR HYBRIDIZING MULTIPLE PROBE PANELS TO NUCLEIC ACID SAMPLES 审中-公开
标题翻译：用于将多个探针面板杂交到核酸样品的方法和工具
公开(公告)号：WO2005033340A2
公开(公告)日：2005-04-14
申请号：PCT/US2004032446
申请日：2004-09-29
申请人： APPLERA CORP , WILLIAMS BRETT , JOHANSEN JACK , HYLDIG-NIELSEN JENS J
发明人： WILLIAMS BRETT , JOHANSEN JACK , HYLDIG-NIELSEN JENS J
IPC分类号： C07H21/04 , C12Q1/68
CPC分类号： C12Q1/6809 , C12Q1/6816 , C12Q1/6883 , C12Q2537/161 , C12Q2537/143 , C12Q2537/1373 , C12Q2565/518
摘要： Described herein are methods and kits that employ multiple probe sets in combination with sequential steps of hybridization analysis for multiplex analysis and/or detection of nucleic acids having one or more distinguishable target sequences.
摘要翻译：本文描述的是使用多个探针组合与杂交分析的顺序步骤进行多重分析和/或检测具有一个或多个可区分靶序列的核酸的方法和试剂盒。

9. 发明申请

WO2004097048A1 PROCESS FOR ASSAY OF NUCLEIC ACID BY COMPETITIVE HYBRIDIZATION USING A DNA MICROARRAY 审中-公开
标题翻译：使用DNA微阵列通过竞争性杂交测定核酸的方法
公开(公告)号：WO2004097048A1
公开(公告)日：2004-11-11
申请号：PCT/JP2004/006214
申请日：2004-04-28
申请人： CANON KABUSHIKI KAISHA , YOSHII, Hiroto , YAMAMOTO, Nobuko , KIMURA, Keiko
发明人： YOSHII, Hiroto , YAMAMOTO, Nobuko , KIMURA, Keiko
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6827 , C12Q1/6837 , C12Q2565/501 , C12Q2527/137 , C12Q2537/161
摘要： A process for rapid and simple assay of nucleic acid on the basis of competitive hybridization using a DNA microarray that comprises a step of hybridizing a labeled nucleic acid other than the target nucleic acid with one of the immobilized probes on the substrate, where the labeled nucleic acid has a known sequence complementary to the probe and can bind thereto specifically.
摘要翻译：基于使用DNA微阵列的竞争性杂交来快速和简单地测定核酸的方法，该DNA微阵列包括将靶核酸以外的标记核酸与底物上的固定化探针之一杂交的步骤，其中标记的核酸酸具有与探针互补的已知序列，并且可以具体结合其。

10. 发明申请

WO2004086041A2 UNIVERSAL READOUT FOR TARGET INDENTIFICATION USING BIOLOGICAL MICROARRAYS 审中-公开
标题翻译：使用生物微阵列进行目标分选的通用读数
公开(公告)号：WO2004086041A2
公开(公告)日：2004-10-07
申请号：PCT/US2004/006539
申请日：2004-03-03
申请人： CORNING INCORPORATED , FANG, Ye
发明人： FANG, Ye
IPC分类号： G01N33/53
CPC分类号： C40B30/04 , B01J2219/00315 , B01J2219/00387 , B01J2219/00497 , B01J2219/00511 , B01J2219/00536 , B01J2219/00576 , B01J2219/00585 , B01J2219/00596 , B01J2219/00605 , B01J2219/0061 , B01J2219/00612 , B01J2219/00621 , B01J2219/00626 , B01J2219/0063 , B01J2219/00637 , B01J2219/00639 , B01J2219/00659 , B01J2219/00677 , B01J2219/00689 , B01J2219/00693 , B01J2219/00722 , B01J2219/00725 , B01J2219/00731 , B01J2219/0074 , B82Y30/00 , C12Q1/6837 , C40B40/06 , C40B40/10 , C40B40/12 , C40B60/14 , G01N2035/00158 , Y10S436/807 , C12Q2537/161
摘要： A method and apparatus for implementing the method is provided. The method involves performing an indirect competitive binding assay on a microarray to identify biological or chemical targets and screen for compounds of interest. The microarray comprises a common ligand located among membrane-, lipid- or protein-associated active binding sites. The method takes advantage of known or well-characterized binding kinetics, and steric interference between biological or chemicals targets of interest and universal readout units for different binding sites within the limited confines of a microspot. The biological targets, chemicals or organisms can specifically bind to target-binding sites, while the universal readout unit binds to the ligands in the microspot.
摘要翻译：提供了用于实现该方法的方法和设备。该方法涉及在微阵列上进行间接竞争结合测定以鉴定生物或化学靶标并筛选感兴趣的化合物。微阵列包含位于膜 - ，脂质或蛋白质相关的活性结合位点中的共同配体。该方法利用已知的或充分表征的结合动力学以及感兴趣的生物或化学目标与用于不同结合位点的通用读出单元之间的空间干扰，所述结合位点位于微点的有限范围内。生物靶标，化学物质或生物体可以特异性结合靶标结合位点，而通用读出单位结合微斑点中的配体。

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