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71. 发明申请

WO2012124965A3 METHOD FOR ASSEMBLING MULTIPLE TARGET LOCI TO A SINGLE NUCLEIC ACID SEQUENCE 审中-公开
标题翻译：将多个目标基因组组装到单核酸序列的方法
公开(公告)号：WO2012124965A3
公开(公告)日：2012-12-27
申请号：PCT/KR2012001808
申请日：2012-03-13
申请人： UNIV YONSEI IACF , BANG DU HEE , HAN HYO JUN
发明人： BANG DU HEE , HAN HYO JUN
IPC分类号： C12Q1/68 , C12N15/11
CPC分类号： C12Q1/686 , C12P19/34 , C12Q1/6858 , C12Q1/6869 , C12Q2531/113 , C12Q2537/143 , C12Q2537/149
摘要： The present invention relates to a method for assembling multiple target loci to a single assembled nucleic acid sequence, and to a method for simultaneously detecting multiple target loci using said assembly method. The method of the present invention involves assembling the multiple target loci to a single assembled nucleic acid sequence through two polymerase chain reactions (PCRs), i.e. a primary PCR and a secondary PCR. More particularly, a pair of primers (a forward primer and an inverse primer) for a primary amplification consist of a target-specific sequence (a target hybridization nucleotide sequence) and a 5'-flanking assembly spacer sequence (an overlapping sequence). In addition, a primary amplified product amplified by means of the pair of primers for a primary amplification is assembled to a single shortened nucleic acid sequence in a convenient and easy manner through a set of primers for a secondary amplification, thus enabling the simultaneous detection of multiple target loci. Accordingly, the method and a kit of the present invention may simultaneously detect and analyze multiple variabilities in DNA sequences of a sample (preferably human blood), thus not only remarkably reducing sequencing costs for detecting variabilities, but also providing a critical approach and means for achieving the concept of customized medicine.
摘要翻译：本发明涉及用于将多个靶基因座组装成单个组装的核酸序列的方法，并且涉及使用所述组装方法同时检测多个靶基因座的方法。本发明的方法包括通过两个聚合酶链式反应（PCR）（即初级PCR和二级PCR）将多个目标基因座组装成单个组装的核酸序列。更具体地说，用于一次扩增的一对引物（正向引物和反向引物）由目标特异性序列（目标杂交核苷酸序列）和5'-侧翼装配间隔区序列（重叠序列）组成。另外，通过用于一次扩增的一对引物将通过用于一次扩增的一对引物扩增的一级扩增产物以方便且容易的方式组装成单一缩短的核酸序列，从而能够同时检测多个目标基因座。因此，本发明的方法和试剂盒可以同时检测和分析样品（优选人血）的DNA序列中的多个变异性，因此不仅显着降低了检测变异性的测序成本，而且还提供了用于实现定制药物的概念。

72. 发明申请

WO2012170433A1 MULTIPLE RNA POLYMERASE TERMINATOR CASSETTES FOR EFFICIENT TRANSCRIPTION OF DISCRETE RNA TEMPLATES 审中-公开
标题翻译：用于有效转录分离RNA模板的多RNA聚合酶终止子CASSETTE
公开(公告)号：WO2012170433A1
公开(公告)日：2012-12-13
申请号：PCT/US2012/040936
申请日：2012-06-05
申请人： SUTRO BIOPHARMA, INC. , MURRAY, Christopher J. , YOUNG, Brian , WILKES, Emily , ROZZELLE, James
发明人： MURRAY, Christopher J. , YOUNG, Brian , WILKES, Emily , ROZZELLE, James
IPC分类号： C12P19/34 , C12N15/10 , C07H21/00
CPC分类号： C12P19/34 , C12N15/63
摘要： This invention provides for an improved and cost efficient means to transcribe circular plasmid DNA into RNA either tRNA or mRNA. The method involves the use of multiple DNA:RNA polymerase termination sequences to prevent waste of high energy phosphates due to inefficient termination of the polymerases.
摘要翻译：本发明提供了将环状质粒DNA转录成tRNA或mRNA的RNA的改进且成本有效的方法。该方法涉及使用多个DNA：RNA聚合酶终止序列，以防止由于聚合酶的低效终止引起的高能量磷酸盐的浪费。

73. 发明申请

WO2012108672A9 REVERSE TRANSCRIPTASE HAVING IMPROVED THERMOSTABILITY 审中-公开
公开(公告)号：WO2012108672A9
公开(公告)日：2012-11-01
申请号：PCT/KR2012000894
申请日：2012-02-07
申请人： BIONEER CORP , PARK HAN OH , YANG SUNG JUN , JOO SUNG MO , HWANG BYOUNG OH
发明人： PARK HAN OH , YANG SUNG JUN , JOO SUNG MO , HWANG BYOUNG OH
IPC分类号： C12N9/12 , C12N15/54 , C12N15/63
CPC分类号： C12N9/1276 , C12N1/20 , C12N9/12 , C12N15/70 , C12P19/34 , Y02P20/52
摘要： The present invention relates to a reverse transcriptase having improved thermostability, more precisely a mutant reverse transcriptase with improved thermostability by substitution of one or more amino acids selected from the group consisting of the 63rd glutamine (Q63), the 264th lysine (K264), the 295th lysine (K295), the 306th threonine (T306), the 346th glutamic acid (E346), the 408th proline (P408), the 438th histidine (H438), and the 454th asparagin (N454) of the amino acid sequence of M-MLV originated reverse transcriptase represented by SEQ. ID. NO: 1 with other amino acids. The mutant reverse transcriptase of the present invention demonstrates excellent thermostability, compared with the wild type reverse transcriptase. Therefore, it is advantageous to obtain the target cDNA with stable reverse transcription activity even in the presence of RNA that can form the stable secondary structure at a high temperature.

74. 发明申请

WO2012124965A2 다중 타겟 위치의 단일 핵산서열로의 어셈블리 방법 审中-公开
标题翻译：用于将多个目标位置组装成单个核酸序列的方法
公开(公告)号：WO2012124965A2
公开(公告)日：2012-09-20
申请号：PCT/KR2012/001808
申请日：2012-03-13
申请人： 연세대학교 산학협력단 , 방두희 , 한효준
发明人： 방두희 , 한효준
IPC分类号： C12Q1/68 , C12N15/11
CPC分类号： C12Q1/686 , C12P19/34 , C12Q1/6858 , C12Q1/6869 , C12Q2531/113 , C12Q2537/143 , C12Q2537/149
摘要： 본 발명은 다중 타겟 위치(multiple target loci)를 하나의 어셈블된 핵산서열로의 어셈블리하는 방법 및 이를 이용한 동시 검출방법에 관한 것이다. 본 발명의 방법은 두 번의 PCR, 즉 제1차 PCR(polymerase chain reaction) 및 제2차 PCR 반응을 통해 다중 타겟 위치를 하나의 어셈블된 핵산서열로 어셈블리시킨다. 보다 상세하게는, 본 발명에서 이용되는 제1차 증폭용 프라이머쌍(정방향 프라이머 및 역방향 프라이머)은 타겟-특이적 서열(타겟 혼성화 뉴클레오타이드 서열) 및 5'-플랭킹 어셈블리 스페이서 서열(오버래핑 서열)로 이루어져 있다. 또한, 상기 제1차 증폭용 프라이머쌍을 이용하여 증폭된 제1차 증폭결과물을 제2차 증폭용 프라이머 세트를 통해 간편하고 용이하게 하나의 단축된 핵산서열로 어셈블리시켜 다중 타겟 위치를 동시에 검출할 수 있다. 따라서, 본 발명의 방법 및 키트는 시료(바람직하게는, 인간의 혈액)의 DNA 서열 상의 다중 변이성(예컨대, SNPs)을 동시에 검출 및 분석함으로써, 변이 검출을 위한 시퀀싱 비용을 현저하게 감소시킬 수 있을 뿐 아니라 맞춤형 의약의 개념을 실현하기 위한 중요한 접근방법 및 수단을 제공한다.
摘要翻译：本发明涉及用于将多个目标基因座组装成单个组装的核酸序列的方法，以及使用所述组装方法同时检测多个靶基因座的方法。本发明的方法包括通过两个聚合酶链反应（PCR）将多个目标基因座组装成单个组装的核酸序列，即初级PCR和二级PCR。更具体地，用于初次扩增的一对引物（正向引物和反向引物）由靶特异性序列（靶杂交核苷酸序列）和5'-侧翼组装间隔物序列（重叠序列）组成。此外，通过用于初级扩增的一对引物扩增的初级扩增产物通过用于二次扩增的一组引物以方便和容易的方式组装成单个缩短的核酸序列，从而能够同时检测多个目标基因座。因此，本发明的方法和试剂盒可以同时检测和分析样品（优选人血液）的DNA序列中的多种变异性，因此不仅显着降低了检测变异性的测序成本，而且提供了关键方法和手段实现定制医学的概念。

75. 发明申请

WO2012097318A2 MODIFIED DNA POLYMERASES FOR IMPROVED AMPLIFICATION 审中-公开
标题翻译：用于改善放大的改性DNA聚合物
公开(公告)号：WO2012097318A2
公开(公告)日：2012-07-19
申请号：PCT/US2012/021348
申请日：2012-01-13
申请人： KAP ABIOSYSTEMS , SCHAFER, Wolfgang , MCEWAN, Paul , VAN DER WALT, Eric , FOSKETT, John , BOURN, William
发明人： SCHAFER, Wolfgang , MCEWAN, Paul , VAN DER WALT, Eric , FOSKETT, John , BOURN, William
IPC分类号： C12N9/12 , C12N15/54 , C12N15/31 , C12N15/63 , C12Q1/48
CPC分类号： C12N9/1252 , C12P19/34 , C12Q1/686 , C12Y207/07007 , Y02P20/52
摘要： The present invention provides improved DNA polymerases that may be better suited for applications in recombinant DNA technologies, in particular technologies involving plant-derived samples. Among other things, the present invention provides modified DNA polymerases derived from directed evolution experiments designed to select mutations that confer advantageous phenotypes under conditions used in industrial or research applications.
摘要翻译：本发明提供了可以更好地适用于重组DNA技术，特别是涉及植物衍生样品的技术的改进的DNA聚合酶。除此之外，本发明提供了衍生自定向进化实验的修饰的DNA聚合酶，设计用于选择在工业或研究应用中使用的条件下赋予有利表型的突变。

76. 发明申请

WO2012092540A1 METHODS FOR THE SYNTHESIS OF POLYADENYLIC ACID 审中-公开
标题翻译：聚亚氨酸合成方法
公开(公告)号：WO2012092540A1
公开(公告)日：2012-07-05
申请号：PCT/US2011/068049
申请日：2011-12-30
申请人： IBIS BIOSCIENCES, INC. , ESHOO, Mark W. , PHILLIPSON, Curtis
发明人： ESHOO, Mark W. , PHILLIPSON, Curtis
IPC分类号： C12P19/34
CPC分类号： C12P19/34 , C12N9/1258 , C12Y207/07008
摘要： The present invention provides compositions, kits, and methods for synthesizing polyadenylic acid using polynucleotide phosphorylase, adenosine diphosphate, a buffering agent, and a divalent metal cation. In certain embodiments, the adenosine diphosphate is present at a concentration between about 5.0 mM and about 100 mM, and the buffering agent has a pH around 8.0. In some embodiments, the prepared polyadenylic acid compositions are free of detectable nucleic acid.
摘要翻译：本发明提供使用多磷酸化酶，二磷酸腺苷，缓冲剂和二价金属阳离子合成聚腺苷酸的组合物，试剂盒和方法。在某些实施方案中，二磷酸腺苷以约5.0mM至约100mM的浓度存在，并且缓冲剂的pH约为8.0。在一些实施方案中，制备的聚腺苷酸组合物不含可检测的核酸。

77. 发明申请

WO2012083072A2 LIGATION METHOD EMPLOYING EUKARYOTIC tRNA LIGASE 审中-公开
标题翻译：使用原核tRNA LIGASE的化学方法
公开(公告)号：WO2012083072A2
公开(公告)日：2012-06-21
申请号：PCT/US2011065268
申请日：2011-12-15
申请人： AGILENT TECHNOLOGIES INC , ZEINER GUSTI , ACH ROBERT A
发明人： ZEINER GUSTI , ACH ROBERT A
IPC分类号： C12N15/10 , C12Q1/25 , C12Q1/68
CPC分类号： C12P19/34 , C12N15/10 , C12N15/1096 , C12Q2521/501 , C12Q2525/191
摘要： Provided herein is a method of preparing an RNA sample comprising: a) obtaining an RNA sample comprising: i. long RNA molecules that may be unfragmented or fragmented to contain 5'-OH group and a 2'-3'-cyclic phosphate group; and ii. short RNA molecules that comprise a 5' phosphate group and a 3' OH group; and b) contacting the RNA sample with an adaptor comprising either a 2'-PO group and 3'-OH group or a 2',3'-cyclic phosphate group in the presence of a eukaryotic tRNA ligase, thereby producing a ligated RNA sample in which a) the short RNA molecules are selectively ligated to the adaptor or b) the short RNA molecules and long RNA fragments are selectively ligated to the adaptor.
摘要翻译：本文提供了制备RNA样品的方法，其包括：a）获得RNA样品，其包含：i。可能未被分裂或片段化以含有5'-OH基团和2'- 3'-环磷酸酯基团的长RNA分子; 和ii。包含5'磷酸基团和3'OH基团的短RNA分子; 和b）在真核tRNA连接酶的存在下，将RNA样品与包含2'-PO基团和3'-OH基团或2'，3'-环状磷酸酯基团的衔接子接触，从而产生连接的RNA样品其中a）将短RNA分子选择性地连接到衔接子上，或b）将短RNA分子和长RNA片段选择性地连接到衔接子上。

78. 发明申请

WO2012055415A1 MICROFLUIDIC DEVICE AND METHOD FOR PROCESSING OF MACROMOLECULES 审中-公开
标题翻译：微流化装置和加工大分子的方法
公开(公告)号：WO2012055415A1
公开(公告)日：2012-05-03
申请号：PCT/DK2011/050402
申请日：2011-10-24
申请人： DANMARKS TEKNISKE UNIVERSITET - DTU , THE CHANCELLOR, MASTERS AND SCHOLARS OF THE UNIVERSITY OF OXFORD , MARIE, Rodolphe , KRISTENSEN, Anders , RASMUSSEN, Kristian Hagsted , MIR, Kalim. Ullah
发明人： MARIE, Rodolphe , KRISTENSEN, Anders , RASMUSSEN, Kristian Hagsted , MIR, Kalim. Ullah
IPC分类号： B01L3/00
CPC分类号： C12P19/34 , B01L3/502746 , B01L3/502761 , B01L2200/06 , B01L2300/0877 , B01L2400/086 , C12Q1/6806 , G01N33/48721 , C12Q2525/204 , C12Q2543/10 , C12Q2565/629
摘要： A microfluidic device and method for enzymatic processing of ultra-long macromolecules is disclosed. The device comprises a reaction chamber with a first manifold, a second manifold, and a plurality of reaction channels, each reaction channel extending from the first manifold to the second manifold. The device further comprises first inlet and outlet channels for filling the reaction channels via the manifolds with one or more macromolecule containers suspended in a first carrier fluid, wherein the first inlet and outlet channels are configured such that a flow established from the first set of inlets to the first set of outlets is guided through the reaction channels, and second inlet and outlet channels for feeding an enzymatic reagent to the reaction chamber essentially without displacing the macromolecule containers trapped in the reaction channels, wherein the second set of inlets and outlets are configured such that a flow established from the second inlet to the second outlet is guided through at least one of the manifolds and bypasses the reaction channels.
摘要翻译：公开了一种用于超长大分子的酶处理的微流体装置和方法。该装置包括具有第一歧管，第二歧管和多个反应通道的反应室，每个反应通道从第一歧管延伸到第二歧管。该装置还包括用于通过歧管用一个或多个大分子容器填充反应通道的第一入口和出口通道，其中悬挂在第一载体流体中，其中第一入口和出口通道被配置成使得从第一组入口第一组出口被引导通过反应通道，第二入口和出口通道用于将酶试剂进料到反应室，基本上不排出被捕获在反应通道中的大分子容器，其中第二组入口和出口被配置使得从第二入口建立到第二出口的流被引导通过歧管中的至少一个并绕过反应通道。

79. 发明申请

WO2012054727A1 REVERSE TRANSCRIPTASE MIXTURES WITH IMPROVED STORAGE STABILITY 审中-公开
标题翻译：具有改进的存储稳定性的反向转录体混合物
公开(公告)号：WO2012054727A1
公开(公告)日：2012-04-26
申请号：PCT/US2011/057103
申请日：2011-10-20
申请人： BIO-RAD LABORATORIES, INC. , MA, Jason , GONG, Xiao-Song
发明人： MA, Jason , GONG, Xiao-Song
IPC分类号： C12P19/34
CPC分类号： C12N9/96 , C12N9/1276 , C12N15/1096 , C12P19/34 , C12Q1/686 , C12Y207/07049 , Y02P20/52
摘要： Reverse transcriptase mixtures with improved storage stability are provided. In one embodiment, a reaction mixture is disclosed comprising sufficient ingredients, except template RNA, for reverse transcription of RNA, the mixture comprising: a reverse transcriptase having an optimal pH above 8; and a buffer, wherein the mixture has a pH of between 6-8, lacks template RNA, and the reverse transcriptase maintains essentially all reverse transcriptase activity following incubation of the mixture at 50° C for 30 minutes.
摘要翻译：提供了具有改进的储存稳定性的逆转录酶混合物。在一个实施方案中，公开了包含用于RNA逆转除的模板RNA之外的足够成分的反应混合物，所述混合物包含：具有最佳pH高于8的逆转录酶; 和缓冲液，其中所述混合物的pH为6-8，缺少模板RNA，并且逆转录酶在50℃温育30分钟后维持基本上所有的逆转录酶活性。

80. 发明申请

WO2012030999A1 AIR COOLING SYSTEMS AND METHODS FOR MICROFLUIDIC DEVICES 审中-公开
标题翻译：空气冷却系统和微流体装置的方法
公开(公告)号：WO2012030999A1
公开(公告)日：2012-03-08
申请号：PCT/US2011/050023
申请日：2011-08-31
申请人： CANON U.S. LIFE SCIENCES, INC. , COURSEY, Johnathan, S. , HASSON, Kenton, C. , LANE, Ben , SCHNEIDER, Eric
发明人： COURSEY, Johnathan, S. , HASSON, Kenton, C. , LANE, Ben , SCHNEIDER, Eric
IPC分类号： F28D7/02
CPC分类号： B01L7/52 , B01L2200/027 , B01L2200/028 , B01L2200/147 , B01L2300/041 , B01L2300/06 , B01L2300/18 , B01L2300/1805 , B01L2300/1838 , B01L2300/1844 , B01L2300/1883 , B01L2300/1894 , C12P19/34 , F28F9/02 , G01N35/08 , G01N2035/00346 , Y02P20/129
摘要： Systems and methods for air cooling a microfluidic device using confinement channels to isolate cooling air from exposed liquids are disclosed. The systems and methods may also thermally condition the cooling airflow for improved robustness of the microfluidic device. In one embodiment, the air cooling system includes a split-level cooling manifold including an inlet duct that directs cooling air to a microfluidic device and an outlet duct that directs air heated by the microfluidic device away from the microfluidic device. The temperature of cooling air may be measured. The cooling air may be preheated to a temperature that is higher than an expected ambient temperature. The temperature of the cooling air after being heated by a microfluidic device may be measured.
摘要翻译：公开了使用限制通道空气冷却微流体装置以将冷却空气与暴露液体隔离的系统和方法。该系统和方法还可以热调节冷却气流，以改善微流体装置的鲁棒性。在一个实施例中，空气冷却系统包括分级冷却歧管，其包括将冷却空气引导至微流体装置的入口管道和将由微流体装置加热的空气远离微流体装置的出口管道。可以测量冷却空气的温度。冷却空气可以被预热到比预期的环境温度高的温度。可以测量由微流体装置加热后的冷却空气的温度。

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