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1.发明授权Amplification method for detection of target nucleic acids involving-fluorescence energy transfer 失效用于检测涉及荧光能量转移的靶核酸的扩增方法公开(公告)号：US07015018B1
公开(公告)日：2006-03-21
申请号：US10048752
申请日：2000-08-03
CPC分类号： C12Q1/6853 , C12Q1/6818 , C12Q2531/113 , C12Q2531/131 , C12Q2565/101
摘要： A method for detecting the presence of a target nucleic acid sequence in a sample, said method comprising subjecting said sample to an amplification reaction using a set of nucleotides, at least one of which is labelled with a first label, and a reagent comprising an amplification primer which can hybridise to said target sequence when in single stranded form and which is connected at its 5′ end to a probe which carries a second label by way of a chemical linking group, said labelled probe being of a sequence which is similar to that of the said target sequence, such that it can hybridise to a complementary region in an amplification product, and wherein one of the first or the second label comprises a donor label and the other comprises an acceptor label, the donor label comprising a fluorescent molecule which is able to donate fluorescent energy to the acceptor label; and monitoring fluorescence of said sample. The method can be used to quantitate the amount of target nucleic acid in the sample as well as to determine sequence characteristics. Kits for effecting the method are also claimed.
摘要翻译： 一种用于检测样品中靶核酸序列的存在的方法，所述方法包括使用一组核苷酸对所述样品进行扩增反应，所述核苷酸中的至少一个用第一标记物标记，并且试剂包含扩增 当单链形式可以与所述靶序列杂交并且其在5'末端连接到通过化学连接基团携带第二标记的探针的引物时，所述标记的探针具有与之相似的序列 的所述靶序列，使得其可以与扩增产物中的互补区域杂交，并且其中所述第一或第二标签中的一个包含供体标记，另一个包含受体标签，所述供体标签包含荧光分子，所述荧光分子 能够将荧光能量捐赠给受体标签; 并监测所述样品的荧光。 该方法可用于定量样品中靶核酸的量以及确定序列特征。 还要求用于实现该方法的套件。
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公开(公告)号：US07264950B1
公开(公告)日：2007-09-04
申请号：US10089498
申请日：2000-09-29
申请人： Martin Alan Lee , Hilary Bird , Dario Lyall Leslie , David James Squirrell , John Shaw , David Wenn , Julie Deacon
发明人： Martin Alan Lee , Hilary Bird , Dario Lyall Leslie , David James Squirrell , John Shaw , David Wenn , Julie Deacon
摘要： A method of carrying out an amplification reaction, said method comprising supplying to a well in a disposable unit (a) a sample which contains or is suspected of containing a target nucleic acid sequence (b) primers, nucleotides and enzymes required to effect said amplification reaction and (c) a buffer system, and subjecting the unit to thermal cycling conditions such that any target nucleic acid present within the sample is amplified; wherein the disposable unit comprises a thermally conducting layer and a facing layer having one or more reagent wells of up to 1000 microns in depth defined therebetween; and the reaction mixture comprises at least one of the following: A) a buffer system wherein the pH is above 8.3; B) a detergent; and/or C) a blocking agent. Apparatus for effecting the method as well as disposable units for use in the method are described. The method is particularly suitable for rapid PCR reactions.
摘要翻译： 一种进行扩增反应的方法，所述方法包括向一次性单元（a）中的井提供含有或怀疑含有靶核酸序列的样品（b）实现所述扩增所需的引物，核苷酸和酶 反应和（c）缓冲系统，并使该单元经受热循环条件，使得存在于样品中的任何靶核酸被扩增; 其中所述一次性单元包括导热层和面对层，其具有在其间限定的深度达1000微米的一个或多个试剂阱; 并且反应混合物包含以下至少一种：A）其中pH高于8.3的缓冲体系; B）洗涤剂; 和/或C）封端剂。 描述了用于实现该方法的装置以及用于该方法的一次性单元。 该方法特别适用于快速PCR反应。
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公开(公告)号：US20090325278A1
公开(公告)日：2009-12-31
申请号：US12553367
申请日：2009-09-03
申请人： Martin Alan Lee , Hilary Bird , Dario Lyall Leslie , David James Squirrell , John Shaw , David Wenn , Julie Deacon
发明人： Martin Alan Lee , Hilary Bird , Dario Lyall Leslie , David James Squirrell , John Shaw , David Wenn , Julie Deacon
IPC分类号： C12M1/34
摘要： A method of carrying out an amplification reaction, said method comprising supplying to a well in a disposable unit (a) a sample which contains or is suspected of containing a target nucleic acid sequence (b) primers, nucleotides and enzymes required to effect said amplification reaction and (c) a buffer system, and subjecting the unit to thermal cycling conditions such that any target nucleic acid present within the sample is amplified; wherein the disposable unit comprises a thermally conducting layer and a facing layer having one or more reagent wells of up to 1000 microns in depth defined therebetween; and the reaction mixture comprises at least one of the following: A) a buffer system wherein the pH is above 8.3; B) a detergent; and/or C) a blocking agent. Apparatus for effecting the method as well as disposable units for use in the method are described. The method is particularly suitable for rapid PCR reactions.
摘要翻译： 一种进行扩增反应的方法，所述方法包括向一次性单元（a）中的井提供含有或怀疑含有靶核酸序列的样品（b）实现所述扩增所需的引物，核苷酸和酶 反应和（c）缓冲系统，并使该单元经受热循环条件，使得存在于样品中的任何靶核酸被扩增; 其中所述一次性单元包括导热层和面对层，其具有在其间限定的深度达1000微米的一个或多个试剂阱; 并且反应混合物包含以下至少一种：A）其中pH高于8.3的缓冲体系; B）洗涤剂; 和/或C）封端剂。 描述了用于实现该方法的装置以及用于该方法的一次性单元。 该方法特别适用于快速PCR反应。
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公开(公告)号：US07659096B2
公开(公告)日：2010-02-09
申请号：US11830283
申请日：2007-07-30
申请人： Martin Alan Lee , Hilary Bird , Dario Lyall Leslie , David James Squirrell , John Shaw , David Wenn , Julie Deacon
发明人： Martin Alan Lee , Hilary Bird , Dario Lyall Leslie , David James Squirrell , John Shaw , David Wenn , Julie Deacon
摘要： A method of carrying out an amplification reaction, said method comprising supplying to a well in a disposable unit (a) a sample which contains or is suspected of containing a target nucleic acid sequence (b) primers, nucleotides and enzymes required to effect said amplification reaction and (c) a buffer system, and subjecting the unit to thermal cycling conditions such that any target nucleic acid present within the sample is amplified; wherein the disposable unit comprises a thermally conducting layer and a facing layer having one or more reagent wells of up to 1000 microns in depth defined therebetween; and the reaction mixture comprises at least one of the following: A) a buffer system wherein the pH is above 8.3; B) a detergent; and/or C) a blocking agent. Apparatus for effecting the method as well as disposable units for use in the method are described. The method is particularly suitable for rapid PCR reactions.
摘要翻译： 一种进行扩增反应的方法，所述方法包括向一次性单元（a）中的井提供含有或怀疑含有靶核酸序列的样品（b）实现所述扩增所需的引物，核苷酸和酶 反应和（c）缓冲系统，并使该单元经受热循环条件，使得存在于样品中的任何靶核酸被扩增; 其中所述一次性单元包括导热层和面对层，其具有在其间限定的深度达1000微米的一个或多个试剂阱; 并且反应混合物包含以下至少一种：A）其中pH高于8.3的缓冲体系; B）洗涤剂; 和/或C）封端剂。 描述了用于实现该方法的装置以及用于该方法的一次性单元。 该方法特别适用于快速PCR反应。
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公开(公告)号：US20080176232A1
公开(公告)日：2008-07-24
申请号：US11830283
申请日：2007-07-30
申请人： Martin Alan Lee , Hilary Bird , Dario Lyall Leslie , David James Squirrell , John Shaw , David Wenn , Julie Deacon
发明人： Martin Alan Lee , Hilary Bird , Dario Lyall Leslie , David James Squirrell , John Shaw , David Wenn , Julie Deacon
摘要： A method of carrying out an amplification reaction, said method comprising supplying to a well in a disposable unit (a) a sample which contains or is suspected of containing a target nucleic acid sequence (b) primers, nucleotides and enzymes required to effect said amplification reaction and (c) a buffer system, and subjecting the unit to thermal cycling conditions such that any target nucleic acid present within the sample is amplified; wherein the disposable unit comprises a thermally conducting layer and a facing layer having one or more reagent wells of up to 1000 microns in depth defined therebetween; and the reaction mixture comprises at least one of the following: A) a buffer system wherein the pH is above 8.3; B) a detergent; and/or C) a blocking agent. Apparatus for effecting the method as well as disposable units for use in the method are described. The method is particularly suitable for rapid PCR reactions.
摘要翻译： 一种进行扩增反应的方法，所述方法包括向一次性单元（a）中的井提供含有或怀疑含有靶核酸序列的样品（b）实现所述扩增所需的引物，核苷酸和酶 反应和（c）缓冲系统，并使该单元经受热循环条件，使得存在于样品中的任何靶核酸被扩增; 其中所述一次性单元包括导热层和面对层，其具有在其间限定的深度达1000微米的一个或多个试剂阱; 并且反应混合物包含以下至少一种：A）其中pH高于8.3的缓冲体系; B）洗涤剂; 和/或C）封端剂。 描述了用于实现该方法的装置以及用于该方法的一次性单元。 该方法特别适用于快速PCR反应。
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公开(公告)号：US08986927B2
公开(公告)日：2015-03-24
申请号：US12553367
申请日：2009-09-03
申请人： Martin Alan Lee , Hilary Bird , Dario Lyall Leslie , David James Squirrell , John Shaw , David Wenn , Julie Deacon
发明人： Martin Alan Lee , Hilary Bird , Dario Lyall Leslie , David James Squirrell , John Shaw , David Wenn , Julie Deacon
摘要： A method of carrying out an amplification reaction, said method comprising supplying to a well in a disposable unit (a) a sample which contains or is suspected of containing a target nucleic acid sequence (b) primers, nucleotides and enzymes required to effect said amplification reaction and (c) a buffer system, and subjecting the unit to thermal cycling conditions such that any target nucleic acid present within the sample is amplified; wherein the disposable unit comprises a thermally conducting layer and a facing layer having one or more reagent wells of up to 1000 microns in depth defined therebetween; and the reaction mixture comprises at least one of the following: A) a buffer system wherein the pH is above 8.3; B) a detergent; and/or C) a blocking agent. Apparatus for effecting the method as well as disposable units for use in the method are described. The method is particularly suitable for rapid PCR reactions.
摘要翻译： 一种进行扩增反应的方法，所述方法包括向一次性单元（a）中的井提供含有或怀疑含有靶核酸序列的样品（b）实现所述扩增所需的引物，核苷酸和酶 反应和（c）缓冲系统，并使该单元经受热循环条件，使得存在于样品中的任何靶核酸被扩增; 其中所述一次性单元包括导热层和面对层，其具有在其间限定的深度达1000微米的一个或多个试剂阱; 并且反应混合物包含以下至少一种：A）其中pH高于8.3的缓冲体系; B）洗涤剂; 和/或C）封端剂。 描述了用于实现该方法的装置以及用于该方法的一次性单元。 该方法特别适用于快速PCR反应。
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公开(公告)号：US20100006453A1
公开(公告)日：2010-01-14
申请号：US12544036
申请日：2009-08-19
CPC分类号： C12Q1/6844 , G01N27/3277 , C12Q2565/607 , C12Q2563/113 , C12Q2561/113
摘要： A method for detecting a target nucleic acid sequence in a sample, by subjecting it to an amplification and taking continuous electrochemical measurements on it during the reaction. The method can be used to determine whet amplification reaction has taken place, to quantitate the amount of target in the sample or to determine sequence characteristics. disclosed is apparatus for use in the method, comprising (i) an amplification reaction vessel which comprises an electrochemic (ii) means for taking continuous electrochemical measurements on a sample contained in the vessel and (iii) tempertature coat measurement means, wherein the electrochemical cell comprises an element formed from an electrically conducting plastics such as a polymer loaded with an electrically conducting material. Further disclosed is a reaction vessel for use in the appar probe for use in the method and a kit for effecting the method.
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公开(公告)号：US06287781B1
公开(公告)日：2001-09-11
申请号：US09622426
申请日：2000-10-19
IPC分类号： C12Q168
CPC分类号： C12Q1/6818 , Y10S435/80 , C12Q2563/107 , C12Q2535/125 , C12Q2531/113
摘要： A method for detecting the presence of a target nucleic acid sequence in a sample is provided. The method comprises subjecting the sample to an amplification reaction to obtain an amplification product where the target nucleic acid sequence is present using a set of nucleotides, at least one of which is fluorescently labelled. The amplification product is contacted with a probe under conditions in which the probe will hybridise to the target sequence. The probe comprises a reactive molecule which is capable of absorbing fluorescence energy from or donating fluorescent energy to the fluorescent labelled nucleotide. The fluorescence of the sample is monitored and related to the presence of the target nucleic acid sequence. The method can be used to quantitate the amount of target nucleic acid in the sample as well as to determine sequence characteristics. Kits for effecting the method are also provided.
摘要翻译： 提供了用于检测样品中靶核酸序列的存在的方法。 该方法包括使样品进行扩增反应以获得使用一组核苷酸存在靶核酸序列的扩增产物，其中至少一个核苷酸被荧光标记。 扩增产物在探针与目标序列杂交的条件下与探针接触。 该探针包含能够从荧光能量吸收荧光能量或向荧光标记的核苷酸提供荧光能量的反应性分子。 监测样品的荧光并与靶核酸序列的存在相关。 该方法可用于定量样品中靶核酸的量以及确定序列特征。 还提供了用于实现该方法的套件。
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