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31. 发明授权

US07312052B1 Polymerase compositions and uses thereof 失效
标题翻译：聚合酶组合物及其用途
公开(公告)号：US07312052B1
公开(公告)日：2007-12-25
申请号：US08529767
申请日：1995-09-18
申请人： Joseph A. Sorge , Rebecca Lynn Mullinax
发明人： Joseph A. Sorge , Rebecca Lynn Mullinax
IPC分类号： C12P19/34 , C12Q1/68 , C07H21/02
CPC分类号： C12Q1/686 , C12Q2521/319 , C12Q2521/101
摘要： The subject invention provides novel compositions containing a mixture of (a) an enzyme that possesses substantial 3′–5′ exonuclease activity (b) a DNA polymerase with less 3′–5′ exonuclease activity than the enzyme with substantial 3′–5′ exonuclease activity. Preferably, the DNA polymerase for inclusion in the compositions are DNA polymerases that substantially lack 3′–5′ exonuclease activity. A preferred embodiment of the invention is a composition comprising the Taq DNA polymerase (isolated from Thermus aquaticus) and the Pfu DNA polymerase (isolated from Pyrococcus furiosus). Another aspect of the invention is to provide methods for synthesizing polynucleotides, typically DNA, using compositions comprising an enzyme that possesses substantial 3′–5′ exonuclease activity and a DNA polymerase with less 3′–5′ exonuclease activity than the enzymes possessing substantial 3′–5′ exonuclease activity, preferably a DNA polymerase that substantially lacks 3′–5′ exonuclease activity. Another aspect of the invention involves the use the subject method of polynucleotide synthesis to carry out the synthesis step in a polymerase chain reaction experiment. Yet another aspect of the invention is to provide kits for the synthesis of polynucleotides, wherein the kits comprise an enzyme that possesses substantial 3′–5′ exonuclease activity and a DNA polymerase with less 3′–5′ exonuclease activity than the enzyme possessing substantial 3′–5′ exonuclease activity.
摘要翻译：本发明提供了包含（a）具有大量3'-5'核酸外切酶活性的酶的混合物的新型组合物（b）具有比实际3'-5'核酸外切酶活性大的3'-5'核酸外切酶活性的3' 核酸外切酶活性。优选地，包含在组合物中的DNA聚合酶是基本上缺乏3'-5'核酸外切酶活性的DNA聚合酶。本发明的优选实施方案是包含Taq DNA聚合酶（从Thermus aquaticus分离）和Pfu DNA聚合酶（从激烈热球菌分离的）的组合物。本发明的另一方面是提供使用包含具有大量3'-5'核酸外切酶活性的酶的组合物和具有比具有实质3的酶的3'-5'外切核酸酶活性少的DNA聚合酶合成多核苷酸（通常为DNA）的方法 '-5'核酸外切酶活性，优选基本上缺乏3'-5'核酸外切酶活性的DNA聚合酶。本发明的另一方面涉及使用聚合酶链反应实验中的多核苷酸合成的主题方法进行合成步骤。本发明的另一方面是提供用于合成多核苷酸的试剂盒，其中所述试剂盒包含具有显着3'-5'核酸外切酶活性的酶和具有比具有实质性的酶的酶更少3'-5'外切核酸酶活性的DNA聚合酶 3'-5'核酸外切酶活性。

32. 发明授权

US06645761B1 Humanized polynucleotide sequence encoding Renilla mulleri green fluorescent protein 有权
标题翻译：人源化多核苷酸序列编码海肾蓝色荧光蛋白
公开(公告)号：US06645761B1
公开(公告)日：2003-11-11
申请号：US09839650
申请日：2001-04-19
申请人： Joseph A. Sorge , Peter Edward Vaillancourt
发明人： Joseph A. Sorge , Peter Edward Vaillancourt
IPC分类号： C12N510
CPC分类号： G01N33/582 , C07K14/43595
摘要： The present invention provides a polynucleotide encoding a green fluorescent protein from Renilla mulleri comprising a humanized sequence which permits enhanced expression of the encoded polypeptide in mammalian cells.
摘要翻译：本发明提供编码来自海肾蓝假单胞菌的绿色荧光蛋白的多核苷酸，其包含允许哺乳动物细胞中编码多肽的增强表达的人源化序列。

33. 发明授权

US06641998B2 Methods and kits to enrich for desired nucleic acid sequences 失效
标题翻译：丰富所需核酸序列的方法和试剂盒
公开(公告)号：US06641998B2
公开(公告)日：2003-11-04
申请号：US09288971
申请日：1999-04-09
申请人： Joseph A. Sorge
发明人： Joseph A. Sorge
IPC分类号： C12Q168
CPC分类号： C12N15/64 , C12N15/1003 , C12N15/66 , C12N15/70
摘要： The invention provides a method and related kits and reagents for producing, purifying, isolating, or enriching desired nucleic acids from a library. The desired nucleic acids are produced from polymerase enzyme extension products, generated from specific primers or sets of primers, in a form that is immediately replicable in a host cell. The invention can be practiced in solution phase, thus eliminating the need for solid phase filter hybridizations, column hybridizations, or gel electrophoresis purification when enriching for or isolating a target sequence or vector from one or more libraries.
摘要翻译：本发明提供了用于从文库产生，纯化，分离或富集所需核酸的方法和相关试剂盒和试剂。所需的核酸由可由宿主细胞中立即可复制的形式的特异性引物或引物组产生的聚合酶延伸产物产生。本发明可以在溶液相中实施，因此当从一个或多个文库富集或分离靶序列或载体时，不需要固相过滤杂交，柱杂交或凝胶电泳纯化。

34. 发明授权

US06589743B2 Methods for detection of a target nucleic acid using a probe comprising secondary structure 有权
公开(公告)号：US06589743B2
公开(公告)日：2003-07-08
申请号：US09981621
申请日：2001-10-17
申请人： Joseph A. Sorge
发明人： Joseph A. Sorge
IPC分类号： C12Q168
CPC分类号： C12Q1/686 , C12Q2565/107 , C12Q2531/125 , C12Q2521/301
摘要： The invention relates to a method of generating a signal indicative of the presence of a target nucleic acid sequence in a sample, where the method includes forming a cleavage structure by incubating a sample comprising a target nucleic acid sequence with a probe having a secondary structure that changes upon binding of the probe to the target nucleic acid sequence, and cleaving the cleavage structure with a nuclease to release a nucleic acid fragment to generate a signal, wherein generation of the signal is indicative of the presence of a target nucleic acid sequence in a sample. The invention also relates to a method of detecting or measuring a target nucleic acid sequence in a sample, where the method includes forming a cleavage structure by incubating a sample containing a target nucleic acid sequence with a probe having a secondary structure that changes upon binding of the probe to a target nucleic acid sequence and cleaving the cleavage structure with a nuclease to generate a cleaved nucleic acid fragment.

35. 发明授权

US5288647A Method of irradiating biological specimens 失效
标题翻译：辐射生物标本的方法
公开(公告)号：US5288647A
公开(公告)日：1994-02-22
申请号：US686491
申请日：1991-04-17
申请人： William C. Zimlich, Jr. , Joseph A. Sorge
发明人： William C. Zimlich, Jr. , Joseph A. Sorge
IPC分类号： C07K17/00 , C12Q1/68 , G01J1/04
CPC分类号： C07K17/00 , C12Q1/68 , Y10T436/25
摘要： A method of irradiating a biological specimen with ultraviolet, in particular a polynucleotide specimen selected from DNA or RNA, or optionally a protein. In the case where the specimen is DNA or RNA, or potentially proteins, the specimen is irradiated to cross-link the specimen to a substrate. In the case where the specimen is DNA, the specimen can also be irradiated to form thymine dimers. The method uses an apparatus which permits relatively precise control of the total ultraviolet dose received by the specimen, despite any changes of ultraviolet flux from the lamps which may occur from during any one experiment, or between a number of experiments. Thus, the method allows relatively highly reproducible results to be obtained.
摘要翻译：用紫外线照射生物样本的方法，特别是选自DNA或RNA的多核苷酸标本，或任选的蛋白质。在标本是DNA或RNA或潜在的蛋白质的情况下，照射样品以将样品交联到基底上。在标本为DNA的情况下，也可以照射样品以形成胸腺嘧啶二聚体。该方法使用允许相对精确地控制由样品接收的总紫外线剂量的装置，尽管在任何一个实验期间或在多个实验之间可能发生来自灯的紫外线通量的任何变化。因此，该方法允许获得相对高度可重复的结果。

36. 发明授权

US5128256A DNA cloning vectors with in vivo excisable plasmids 失效
标题翻译：具有体内可切除质粒的DNA克隆载体
公开(公告)号：US5128256A
公开(公告)日：1992-07-07
申请号：US341261
申请日：1989-04-20
申请人： William Huse , Joseph A. Sorge , Jay M. Short
发明人： William Huse , Joseph A. Sorge , Jay M. Short
IPC分类号： C12N15/64 , C12N15/70
CPC分类号： C12N15/70 , C12N15/64
摘要： Vectors are described that circumvent traditional DNA cloning and subcloning procedures, and that contain a unique DNA cartridge that permits both cloning of DNA directly into DNA sequences present within the cartridge, and in vivo removal and circularization of the cartridge thereby yielding an autonomously replicating structure. Because the DNA cartridge can include a wide variety of functional DNA sequences, the cloned DNA can be subjected to a plethora of molecular biological procedures without having to remove the cloned DNA from the cartridge thereby obviating the need to perform additional subcloning techniques. A particularly useful example of this type of vector is bacteriophage lambda containing the DNA cartridge.
摘要翻译：描述了可以克服传统DNA克隆和亚克隆程序的载体，并且其含有独特的DNA柱，其允许将DNA直接克隆到存在于盒内的DNA序列中，并且在体内移除和环化圆筒，从而产生自主复制的结构。因为DNA盒可以包括多种功能性DNA序列，所以克隆的DNA可以经受大量的分子生物学过程，而不必从盒中除去克隆的DNA，从而避免进行附加亚克隆技术的需要。这种载体的特别有用的实例是含有DNA盒的噬菌体λ。

37. 发明授权

US08314220B2 Methods compositions, and kits for detection of microRNA 有权
标题翻译：用于检测microRNA的方法，组合物和试剂盒
公开(公告)号：US08314220B2
公开(公告)日：2012-11-20
申请号：US11627926
申请日：2007-01-26
申请人： Rebecca L. Mullinax , Joseph A. Sorge
发明人： Rebecca L. Mullinax , Joseph A. Sorge
IPC分类号： C07H21/02 , C12Q1/68 , C07H21/04
CPC分类号： C12Q1/6844 , C12Q1/6811 , C12Q1/6858 , Y10T436/143333 , C12Q2525/207
摘要： The present invention provides methods, nucleic acids, compositions, and kits for detecting microRNA (miRNA) in samples. The methods comprise designing mRNA-specific primers, adding a polyA tail to the miRNA, and using reverse transcription and amplification to detect the miRNA. The nucleic acids, compositions, and kits typically comprise some or all of the components necessary to practice the methods of the invention.
摘要翻译：本发明提供了用于在样品中检测微小RNA（miRNA）的方法，核酸，组合物和试剂盒。该方法包括设计mRNA特异性引物，向miRNA中加入polyA尾，并使用逆转录和扩增检测miRNA。核酸，组合物和试剂盒通常包含实施本发明方法所需的部分或全部组分。

38. 发明授权

US07932070B2 High fidelity DNA polymerase compositions and uses therefor 有权
标题翻译：高保真DNA聚合酶组合物及其用途
公开(公告)号：US07932070B2
公开(公告)日：2011-04-26
申请号：US10227110
申请日：2002-08-23
申请人： Holly Hogrefe , Michael Borns , Joseph A. Sorge
发明人： Holly Hogrefe , Michael Borns , Joseph A. Sorge
IPC分类号： C12N9/12 , C07K1/00
CPC分类号： C12N9/1252
摘要： The subject invention relates to compositions comprising an enzyme mixture which comprises a first enzyme and a second enzyme, where the first enzyme comprises a DNA polymerization activity and the second enzyme comprises an 3′-5′ exonuclease activity and a reduced DNA polymerization activity. The invention also relates to the above compositions in kit format and methods for high fidelity DNA synthesis using the subject compositions of the invention.
摘要翻译：本发明涉及包含含有第一酶和第二酶的酶混合物的组合物，其中第一种酶包含DNA聚合活性，第二种酶包含3'-5'核酸外切酶活性和降低的DNA聚合活性。本发明还涉及使用本发明的主题组合物的用于高保真DNA合成的试剂盒形式的上述组合物和方法。

39. 发明授权

US07824859B2 Methods for detection of a target nucleic acid by forming a cleavage structure using an RNA polymerase 有权
标题翻译：通过使用RNA聚合酶形成切割结构来检测靶核酸的方法
公开(公告)号：US07824859B2
公开(公告)日：2010-11-02
申请号：US11476893
申请日：2006-06-22
申请人： Joseph A. Sorge
发明人： Joseph A. Sorge
IPC分类号： C12Q1/68 , C07H21/02 , C07H21/04
CPC分类号： G01N33/5308 , C12Q1/48 , C12Q1/6823 , C12Q2521/119 , C12Q2521/301 , C12Q2531/113 , C12Q2521/307 , C12Q2525/301 , C12Q2565/519 , C12Q2561/109 , C12Q2525/161
摘要： The invention relates to compositions and methods for generating a signal indicative of the presence of a target nucleic acid in a sample, where the compositions and methods include an RNA polymerase, a FEN nuclease, and a probe.
摘要翻译：本发明涉及用于产生指示样品中目标核酸存在的信号的组合物和方法，其中组合物和方法包括RNA聚合酶，FEN核酸酶和探针。

40. 发明授权

US07736854B2 Methods of detection of a target nucleic acid sequence 有权
标题翻译：检测靶核酸序列的方法
公开(公告)号：US07736854B2
公开(公告)日：2010-06-15
申请号：US11606518
申请日：2006-11-29
申请人： Joseph A. Sorge
发明人： Joseph A. Sorge
IPC分类号： C12Q1/68 , C07H21/02 , C07H21/04
CPC分类号： C12Q1/6823 , C12Q1/6816 , C12Q2521/301 , C12Q2531/113 , C12Q2521/307 , C12Q2525/161 , C12Q2525/301 , C12Q2561/109 , C12Q2565/519 , C12Q2561/101 , C12Q2521/107
摘要： The present invention is directed to methods of generating a signal indicative of the presence of a target nucleic acid in a sample by forming a cleavage structure comprising duplex and single-stranded nucleic acid by incubating a sample comprising a target nucleic acid with a thermostable nucleic acid polymerase substantially lacking 5′ to 3′ exonuclease activity and cleaving said cleavage structure with a thermostable Fen nuclease lacking 5′ to 3′ synthetic activity to generate a signal.
摘要翻译：本发明涉及通过形成包含双链体和单链核酸的切割结构，通过将包含靶核酸的样品与热稳定性核酸一起孵育来产生指示样品中目标核酸存在的信号的方法聚合酶基本上缺乏5'至3'核酸外切酶活性，并用缺乏5'至3'合成活性的热稳定的Fen核酸酶切割所述切割结构以产生信号。

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