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1. 发明申请

US20050037361A1 Methods for the reduction of stutter in microsatellite amplification 有权
公开(公告)号：US20050037361A1
公开(公告)日：2005-02-17
申请号：US10637466
申请日：2003-08-08
申请人： Sulekha Rao Coticone , Will Bloch
发明人： Sulekha Rao Coticone , Will Bloch
IPC分类号： G01N33/53 , C12N15/09 , C12Q1/68 , G01N33/566 , C12P19/34
CPC分类号： C12Q1/686 , C12Q1/6846 , C12Q1/6848 , C12Q1/6876 , C12Q2527/125 , C12Q2525/151
摘要： The invention provides a method for reducing stutter in the amplification of a microsatellite comprising the steps of providing a sample comprising a microsatellite having a G+C content of 50% or less; contacting the sample with at least one enzyme having nucleic acid polymerase activity; and incubating the sample with the enzyme for a sufficient amount of time and under conditions sufficient to amplify the microsatellite; wherein the incubation is performed in the presence of an amount of betaine, sorbitol or mixtures thereof, effective to reduce stutter relative to the amount of stutter observed in the absence of betaine and/or sorbitol. The invention also provides compositions containing betaine and/or sorbitol, kits for amplifying microsatellites having a G+C content of 50% or less, and methods of using all of the foregoing.

2. 发明授权

US06841349B2 Methods for the reduction of stutter in microsatellite amplification 有权
标题翻译：减少微卫星放大的口吃的方法
公开(公告)号：US06841349B2
公开(公告)日：2005-01-11
申请号：US09850514
申请日：2001-05-07
申请人： Sulekha Rao Coticone , Will Bloch
发明人： Sulekha Rao Coticone , Will Bloch
IPC分类号： G01N33/53 , C12N15/09 , C12Q1/68 , G01N33/566 , C07H21/02 , C07H21/04
CPC分类号： C12Q1/686 , C12Q1/6846 , C12Q1/6848 , C12Q1/6876 , C12Q2527/125 , C12Q2525/151
摘要： The invention provides a method for reducing stutter in the amplification of a microsatellite comprising the steps of providing a sample comprising a microsatellite having a G+C content of 50% or less; contacting the sample with at least one enzyme having nucleic acid polymerase activity; and incubating the sample with the enzyme for a sufficient amount of time and under conditions sufficient to amplify the microsatellite; wherein the incubation is performed in the presence of an amount of betaine, sorbitol or mixtures thereof, effective to reduce stutter relative to the amount of stutter observed in the absence of betaine and/or sorbitol. The invention also provides compositions containing betaine and/or sorbitol, kits for amplifying microsatellites having a G+C content of 50% or less, and methods of using all of the foregoing.
摘要翻译：本发明提供了减少微卫星扩增中的口吃的方法，包括以下步骤：提供包含G + C含量为50％或更少的微卫星的样品; 使样品与至少一种具有核酸聚合酶活性的酶接触; 并将样品与酶温育足够的时间和足以扩增微卫星的条件; 其中在一定量的甜菜碱，山梨糖醇或其混合物的存在下进行培养，其有效地相对于在不存在甜菜碱和/或山梨醇的情况下观察到的口吃量减少口吃。本发明还提供含有甜菜碱和/或山梨糖醇的组合物，用于扩增G + C含量为50％以下的微卫星的试剂盒，以及使用上述所有方法。

3. 发明申请

US20080213859A1 Methods for the Reduction of Stutter in Microsatellite Amplification 审中-公开
标题翻译：减少微卫星放大技术的方法
公开(公告)号：US20080213859A1
公开(公告)日：2008-09-04
申请号：US11742524
申请日：2007-04-30
申请人： Sulekha Rao Coticone , Will Bloch
发明人： Sulekha Rao Coticone , Will Bloch
IPC分类号： C12N9/10
CPC分类号： C12Q1/686 , C12Q1/6846 , C12Q1/6848 , C12Q1/6876 , C12Q2527/125 , C12Q2525/151
摘要： The invention provides a method for reducing stutter in the amplification of a microsatellite comprising the steps of providing a sample comprising a microsatellite having a G+C content of 50% or less; contacting the sample with at least one enzyme having nucleic acid polymerase activity; and incubating the sample with the enzyme for a sufficient amount of time and under conditions sufficient to amplify the microsatellite; wherein the incubation is performed in the presence of an amount of betaine, sorbitol or mixtures thereof, effective to reduce stutter relative to the amount of stutter observed in the absence of betaine and/or sorbitol. The invention also provides compositions containing betaine and/or sorbitol, kits for amplifying microsatellites having a G+C content of 50% or less, and methods of using all of the foregoing.
摘要翻译：本发明提供了减少微卫星扩增中的口吃的方法，包括以下步骤：提供包含G + C含量为50％或更少的微卫星的样品; 使样品与至少一种具有核酸聚合酶活性的酶接触; 并将样品与酶温育足够的时间和足以扩增微卫星的条件; 其中在一定量的甜菜碱，山梨糖醇或其混合物的存在下进行培养，其有效地相对于在不存在甜菜碱和/或山梨醇的情况下观察到的口吃量减少口吃。本发明还提供含有甜菜碱和/或山梨糖醇的组合物，用于扩增G + C含量为50％以下的微卫星的试剂盒，以及使用上述所有方法。

4. 发明授权

US06783940B2 Method of reducing non-specific amplification in PCR 有权
标题翻译：降低PCR中非特异性扩增的方法
公开(公告)号：US06783940B2
公开(公告)日：2004-08-31
申请号：US09998887
申请日：2001-10-31
申请人： Ian J. McLaughlin , Sulekha Rao Coticone , Will Bloch
发明人： Ian J. McLaughlin , Sulekha Rao Coticone , Will Bloch
IPC分类号： C12Q168
CPC分类号： C12Q1/686 , C12Q2527/125
摘要： The invention provides methods for reducing non-specific amplification DNA in a polymerase chain reaction comprising providing a sample comprising a target DNA sequence of interest; contacting the sample with at least one enzyme having nucleic acid polymerase activity; and incubating the sample with said enzyme for a time and under conditions sufficient to amplify the target DNA sequence, forming amplified target sequence; wherein the incubation is performed in the presence of an amount of sorbitol, or sorbitol and DMSO effective to reduce the non-specific amplification relative to the amount of non-specific amplification observed in the absence of sorbitol, or sorbitol and DMSO. The methods are suitable for amplification of ribosomal DNA, particularly from clinical samples. Compositions and kits containing sorbitol, or sorbitol and DMSO for reducing non-specific amplification are also provided.
摘要翻译：本发明提供了降低聚合酶链反应中的非特异性扩增DNA的方法，包括提供包含目标DNA序列的样品; 使样品与至少一种具有核酸聚合酶活性的酶接触; 并将样品与所述酶一起孵育一段时间并在足以扩增靶DNA序列的条件下形成扩增的靶序列; 其中在一定量的山梨糖醇或山梨糖醇和DMSO的存在下进行培养，所述山梨糖醇或山梨糖醇和DMSO相对于在不存在山梨醇或山梨糖醇和DMSO的情况下观察到的非特异性扩增量有效减少非特异性扩增。该方法适用于扩增核糖体DNA，特别是临床样品。还提供了含有山梨醇或山梨糖醇和DMSO用于降低非特异性扩增的组合物和试剂盒。

5. 发明授权

US07211385B2 Methods for the reduction of stutter microsatellite amplification 有权
标题翻译：减少迷路微卫星扩增的方法
公开(公告)号：US07211385B2
公开(公告)日：2007-05-01
申请号：US10637466
申请日：2003-08-08
申请人： Sulekha Rao Coticone , Will Bloch
发明人： Sulekha Rao Coticone , Will Bloch
IPC分类号： C12Q1/68 , C12P19/34 , C07H21/02 , C07H21/04
CPC分类号： C12Q1/686 , C12Q1/6846 , C12Q1/6848 , C12Q1/6876 , C12Q2527/125 , C12Q2525/151
摘要： The invention provides a method for reducing stutter in the amplification of a microsatellite comprising the steps of providing a sample comprising a microsatellite having a G+C content of 50% or less; contacting the sample with at least one enzyme having nucleic acid polymerase activity; and incubating the sample with the enzyme for a sufficient amount of time and under conditions sufficient to amplify the microsatellite; wherein the incubation is performed in the presence of an amount of betaine, sorbitol or mixtures thereof, effective to reduce stutter relative to the amount of stutter observed in the absence of betaine and/or sorbitol. The invention also provides compositions containing betaine and/or sorbitol, kits for amplifying microsatellites having a G+C content of 50% or less, and methods of using all of the foregoing.
摘要翻译：本发明提供了减少微卫星扩增中的口吃的方法，包括以下步骤：提供包含G + C含量为50％或更少的微卫星的样品; 使样品与至少一种具有核酸聚合酶活性的酶接触; 并将样品与酶温育足够的时间和足以扩增微卫星的条件; 其中在一定量的甜菜碱，山梨糖醇或其混合物的存在下进行培养，其有效地相对于在不存在甜菜碱和/或山梨醇的情况下观察到的口吃量减少口吃。本发明还提供含有甜菜碱和/或山梨糖醇的组合物，用于扩增G + C含量为50％以下的微卫星的试剂盒，以及使用上述所有方法。

6. 发明授权

US06780588B2 Methods for the reduction of stutter in microsatellite amplification 有权
标题翻译：减少微卫星放大的口吃的方法
公开(公告)号：US06780588B2
公开(公告)日：2004-08-24
申请号：US09850590
申请日：2001-05-07
申请人： Sulekha Rao Coticone , Will Bloch
发明人： Sulekha Rao Coticone , Will Bloch
IPC分类号： C12Q168
CPC分类号： C12Q1/6848 , C12Q2549/125 , C12Q2527/137 , C12Q2521/101
摘要： The invention provides a method for reducing stutter in the amplification of a microsatellite comprising the steps of providing a sample comprising a microsatellite having a G+C content of greater than 50%; contacting the sample with at least one enzyme having nucleic acid polymerase activity; and incubating the sample with the enzyme for a sufficient amount of time and under conditions sufficient to amplify the microsatellite; wherein the incubation is performed in the presence of an amount of sorbitol effective to reduce stutter relative to the amount of stutter observed in the absence of sorbitol. The invention also provides compositions containing sorbitol, kits for amplifying microsatellites having a G+C content of greater than 50%, and methods of using all of the foregoing.
摘要翻译：本发明提供了减少微卫星扩增中的口吃的方法，包括提供包含G + C含量大于50％的微卫星样品的步骤; 使样品与至少一种具有核酸聚合酶活性的酶接触; 并将样品与酶温育足够的时间和足以扩增微卫星的条件; 其中在一定量的山梨糖醇的存在下进行培养，其有效地减少相对于在不存在山梨糖醇时观察到的口吃量的口吃。本发明还提供含有山梨醇的组合物，用于扩增G + C含量大于50％的微卫星的试剂盒，以及使用上述所有方法。

7. 发明申请

US20130011839A1 Methods for the Reduction of Stutter in Microsatellite Amplification 有权
公开(公告)号：US20130011839A1
公开(公告)日：2013-01-10
申请号：US13611974
申请日：2012-09-12
申请人： Sulekha Rao Coticone , Will Bloch
发明人： Sulekha Rao Coticone , Will Bloch
IPC分类号： C12Q1/68 , C12N9/12 , C12P19/34
CPC分类号： C12Q1/686 , C12Q1/6846 , C12Q1/6848 , C12Q1/6876 , C12Q2527/125 , C12Q2525/151
摘要： The invention provides a method for reducing stutter in the amplification of a microsatellite comprising the steps of providing a sample comprising a microsatellite having a G+C content of 50% or less; contacting the sample with at least one enzyme having nucleic acid polymerase activity; and incubating the sample with the enzyme for a sufficient amount of time and under conditions sufficient to amplify the microsatellite; wherein the incubation is performed in the presence of an amount of betaine, sorbitol or mixtures thereof, effective to reduce stutter relative to the amount of stutter observed in the absence of betaine and/or sorbitol. The invention also provides compositions containing betaine and/or sorbitol, kits for amplifying microsatellites having a G+C content of 50% or less, and methods of using all of the foregoing.

8. 发明申请

US20110143352A1 Methods for the Reduction of Stutter in Microsatellite Amplification 审中-公开
标题翻译：减少微卫星放大技术的方法
公开(公告)号：US20110143352A1
公开(公告)日：2011-06-16
申请号：US12769577
申请日：2010-04-28
申请人： Sulekha Rao COTICONE , Will Bloch
发明人： Sulekha Rao COTICONE , Will Bloch
IPC分类号： C12Q1/68 , C12N9/12
CPC分类号： C12Q1/686 , C12Q1/6846 , C12Q1/6848 , C12Q1/6876 , C12Q2527/125 , C12Q2525/151
摘要： The invention provides a method for reducing stutter in the amplification of a microsatellite comprising the steps of providing a sample comprising a microsatellite having a G+C content of 50% or less; contacting the sample with at least one enzyme having nucleic acid polymerase activity; and incubating the sample with the enzyme for a sufficient amount of time and under conditions sufficient to amplify the microsatellite; wherein the incubation is performed in the presence of an amount of betaine, sorbitol or mixtures thereof, effective to reduce stutter relative to the amount of stutter observed in the absence of betaine and/or sorbitol. The invention also provides compositions containing betaine and/or sorbitol, kits for amplifying microsatellites having a G+C content of 50% or less, and methods of using all of the foregoing.
摘要翻译：本发明提供了减少微卫星扩增中的口吃的方法，包括以下步骤：提供包含G + C含量为50％或更少的微卫星的样品; 使样品与至少一种具有核酸聚合酶活性的酶接触; 并将样品与酶温育足够的时间和足以扩增微卫星的条件; 其中在一定量的甜菜碱，山梨糖醇或其混合物的存在下进行培养，其有效地相对于在不存在甜菜碱和/或山梨醇的情况下观察到的口吃量减少口吃。本发明还提供含有甜菜碱和/或山梨糖醇的组合物，用于扩增G + C含量为50％以下的微卫星的试剂盒，以及使用上述所有方法。

9. 发明授权

US08628925B2 Linear amplification of short nucleic acids 有权
标题翻译：短核酸的线性扩增
公开(公告)号：US08628925B2
公开(公告)日：2014-01-14
申请号：US13154128
申请日：2011-06-06
申请人： Will Bloch
发明人： Will Bloch
IPC分类号： C12Q1/68 , C12P19/34
CPC分类号： C12P19/34 , C12Q1/6844 , C12Q1/6853 , C12Q2531/101 , C12Q2525/301 , C12Q2521/107 , C12Q2527/125
摘要： The present teachings provide novel methods for amplifying short nucleic acids. In some embodiments, the present teachings provide novel methods for linearly amplifying a collection of micro RNAs by using temperature cycling during a reverse transcription reaction. The cycling can comprise at least 20 cycles of an annealing temperature segment of 10° C.-30° C., and a denaturation temperature segment of 35° C.-60° C. In some embodiments, the temperature cycled reaction can comprise an osmolyte.
摘要翻译：本教导提供了用于扩增短核酸的新方法。在一些实施方案中，本发明提供了用于在逆转录反应期间通过使用温度循环线性扩增微RNA集合的新方法。循环可以包括10℃-30℃的退火温度段和35℃-60℃的变性温度段的至少20个循环。在一些实施方案中，温度循环反应可以包括渗透剂

10. 发明授权

US5538871A In situ polymerase chain reaction 失效
标题翻译：原位聚合酶链反应
公开(公告)号：US5538871A
公开(公告)日：1996-07-23
申请号：US390256
申请日：1995-02-17
申请人： Gerard J. Nuovo , Will Bloch
发明人： Gerard J. Nuovo , Will Bloch
IPC分类号： A61B10/00 , B01L7/00 , C12M1/40 , C12N15/09 , C12Q1/68 , C12R1/19 , C12P19/34 , C07H21/04 , C12N15/00
CPC分类号： C12Q1/6841 , B01L7/52 , C12Q1/686
摘要： Improvements to the in situ polymerase chain reaction (PCR), a process of in vitro enzymatic amplification of specific nucleic acid sequences within the cells where they originate, can be achieved by changing the way that the enzymatic reaction is started. Reaction initiation is delayed until the start of PCR thermal cycling, either by withholding a subset of PCR reagents from the cellular preparation until the preparation has been heated to 50.degree. C. to 80.degree. C., immediately before thermal cycling is begun, or by adding to the PCR reagents a single-stranded DNA binding protein which blocks reaction at temperatures below about 50.degree. C. If the in situ PCR is performed on cellular preparations already attached to a microscope slide, thermal cycling also is facilitated by use of a thermal cycler sample block or compartment designed optimally to hold the microscope slide and any vapor barrier covering the slide.
摘要翻译：原位聚合酶链反应（PCR）的改进，其可以通过改变酶反应开始的方式来实现其在其中起源的细胞内的特定核酸序列的体外酶促扩增过程。反应开始被延迟直到PCR热循环的开始，通过从细胞制剂中扣留一组PCR试剂，直到制备在热循环开始之前立即加热至50℃至80℃，或通过向PCR试剂中加入在低于约50℃的温度下阻断反应的单链DNA结合蛋白。如果在已经连接到显微镜载玻片上的细胞制剂上进行原位PCR，则通过使用热循环仪样品块或隔室设计最佳，以保持显微镜载玻片和覆盖幻灯片的任何蒸汽屏障。

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