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公开(公告)号：US10329603B2
公开(公告)日：2019-06-25
申请号：US15496756
申请日：2017-04-25
申请人： Alere San Diego Inc.
IPC分类号： C12P19/34 , C12Q1/6844 , C12Q1/6806 , C12N9/12 , B01L7/00 , C12Q1/6818 , G01N33/53
摘要： This disclosure describes related novel methods for Recombinase-Polymerase Amplification (RPA) of a target DNA that exploit the properties of recombinase and related proteins, to invade double-stranded DNA with single stranded homologous DNA permitting sequence specific priming of DNA polymerase reactions. The disclosed methods have the advantage of not requiring thermocycling or thermophilic enzymes, thus offering easy and affordable implementation and portability relative to other amplification methods. Further disclosed are conditions to enable real-time monitoring of RPA reactions, methods to regulate RPA reactions using light and otherwise, methods to determine the nature of amplified species without a need for gel electrophoresis, methods to improve and optimize signal to noise ratios in RPA reactions, methods to optimize oligonucleotide primer function, methods to control carry-over contamination, and methods to employ sequence-specific third ‘specificity’ probes. Further described are novel properties and approaches for use of probes monitored by light in dynamic recombination environments.
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公开(公告)号：US10329602B2
公开(公告)日：2019-06-25
申请号：US15395446
申请日：2016-12-30
IPC分类号： C12P19/34 , C12Q1/6844 , C12Q1/6848 , C12Q1/686 , C12Q1/70
摘要： This disclosure describes related novel methods for Recombinase-Polymerase Amplification (RPA) of a target DNA that exploit the properties of recombinase and related proteins, to invade double-stranded DNA with single stranded homologous DNA permitting sequence specific priming of DNA polymerase reactions. The disclosed methods have the advantage of not requiring thermocycling or thermophilic enzymes. Further, the improved processivity of the disclosed methods may allow amplification of DNA up to hundres of megabases in length.
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公开(公告)号：US20190136300A1
公开(公告)日：2019-05-09
申请号：US16079273
申请日：2017-02-24
申请人： ALERE SAN DIEGO INC
发明人： Frank Ray Bowler , Grzegorz Artur Orlowski , Hazel Lucy Greetham , Cheng Zhou , Niall A. Armes , Olaf Piepenburg
IPC分类号： C12Q1/6825 , C12Q1/6876
摘要： This invention relates to sequence specific electrochemically-labeled oligonucleotide probes for the detection of nucleic acids and methods associated therewith.
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公开(公告)号：US20170321262A1
公开(公告)日：2017-11-09
申请号：US15496756
申请日：2017-04-25
申请人： Alere San Diego Inc.
CPC分类号： C12Q1/6844 , B01L7/52 , B01L2200/10 , B01L2300/0627 , B01L2300/18 , C12N9/1252 , C12Q1/6806 , C12Q1/6818 , C12Y207/07007 , G01N33/5308 , C12Q2531/113 , C12Q2522/101 , C12Q2521/507 , C12Q2527/125
摘要： This disclosure describes related novel methods for Recombinase-Polymerase Amplification (RPA) of a target DNA that exploit the properties of recombinase and related proteins, to invade double-stranded DNA with single stranded homologous DNA permitting sequence specific priming of DNA polymerase reactions. The disclosed methods have the advantage of not requiring thermocycling or thermophilic enzymes, thus offering easy and affordable implementation and portability relative to other amplification methods. Further disclosed are conditions to enable real-time monitoring of RPA reactions, methods to regulate RPA reactions using light and otherwise, methods to determine the nature of amplified species without a need for gel electrophoresis, methods to improve and optimize signal to noise ratios in RPA reactions, methods to optimize oligonucleotide primer function, methods to control carry-over contamination, and methods to employ sequence-specific third ‘specificity’ probes. Further described are novel properties and approaches for use of probes monitored by light in dynamic recombination environments.
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公开(公告)号：US09663820B2
公开(公告)日：2017-05-30
申请号：US14989499
申请日：2016-01-06
申请人： Alere San Diego Inc.
CPC分类号： C12Q1/6844 , B01L7/52 , B01L2200/10 , B01L2300/0627 , B01L2300/18 , C12N9/1252 , C12Q1/6806 , C12Q1/6818 , C12Y207/07007 , G01N33/5308 , C12Q2531/113 , C12Q2522/101 , C12Q2521/507 , C12Q2527/125
摘要： This disclosure describes related novel methods for Recombinase-Polymerase Amplification (RPA) of a target DNA that exploit the properties of recombinase and related proteins, to invade double-stranded DNA with single stranded homologous DNA permitting sequence specific priming of DNA polymerase reactions. The disclosed methods have the advantage of not requiring thermocycling or thermophilic enzymes, thus offering easy and affordable implementation and portability relative to other amplification methods. Further disclosed are conditions to enable real-time monitoring of RPA reactions, methods to regulate RPA reactions using light and otherwise, methods to determine the nature of amplified species without a need for gel electrophoresis, methods to improve and optimize signal to noise ratios in RPA reactions, methods to optimize oligonucleotide primer function, methods to control carry-over contamination, and methods to employ sequence-specific third ‘specificity’ probes. Further described are novel properties and approaches for use of probes monitored by light in dynamic recombination environments.
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公开(公告)号：US20160083780A1
公开(公告)日：2016-03-24
申请号：US14865707
申请日：2015-09-25
CPC分类号： C12Q1/6844 , B01L7/52 , B01L2200/10 , B01L2300/0627 , B01L2300/18 , C12N9/1252 , C12Q1/6806 , C12Q1/6818 , C12Y207/07007 , G01N33/5308 , C12Q2531/113 , C12Q2522/101 , C12Q2521/507 , C12Q2527/125
摘要： This disclosure describes related novel methods for Recombinase-Polymerase Amplification (RPA) of a target DNA that exploit the properties of recombinase and related proteins, to invade double-stranded DNA with single stranded homologous DNA permitting sequence specific priming of DNA polymerase reactions. The disclosed methods have the advantage of not requiring thermocycling or thermophilic enzymes, thus offering easy and affordable implementation and portability relative to other amplification methods. Further disclosed are conditions to enable real-time monitoring of RPA reactions, methods to regulate RPA reactions using light and otherwise, methods to determine the nature of amplified species without a need for gel electrophoresis, methods to improve and optimize signal to noise ratios in RPA reactions, methods to optimize oligonucleotide primer function, methods to control carry-over contamination, and methods to employ sequence-specific third ‘specificity’ probes. Further described are novel properties and approaches for use of probes monitored by light in dynamic recombination environments.
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公开(公告)号：US10093908B2
公开(公告)日：2018-10-09
申请号：US14135227
申请日：2013-12-19
IPC分类号： C07K14/00 , C12N9/12 , C12N9/00 , C12Q1/6844
摘要： The present invention features novel, diverse, hybrid and engineered recombinase enzymes, and the utility of such proteins with associated recombination factors for carrying out DNA amplification assays. The present invention also features different recombinase ‘systems’ having distinct biochemical activities in DNA amplification assays, and differing requirements for loading factors, single-stranded DNA binding proteins (SSBs), and the quantity of crowding agent employed.
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公开(公告)号：US20170175173A1
公开(公告)日：2017-06-22
申请号：US15284065
申请日：2016-10-03
申请人： Alere San Diego Inc.
发明人： Olaf Piepenburg , Niall A. Armes
CPC分类号： C12Q1/6816 , C07H21/00 , C12Q2521/301 , C12Q2521/531 , C12Q2525/119
摘要： A new class of nucleic acid substrates for AP endonucleases and members of the glycosylase/lyase family of enzymes is described. Representatives of each family, the enzymes Nfo and fpg, respectively, cleave nucleic acid backbones at positions in which a base has been replaced by a linker to which a variety of label moieties may be attached. The use of these synthetic substrates embedded within oligonucleotides is of utility in a number of applications.
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公开(公告)号：US20160326502A1
公开(公告)日：2016-11-10
申请号：US14135227
申请日：2013-12-19
CPC分类号： C12N9/1241 , C12N9/00 , C12Q1/6844 , C12Y207/07 , C12Q2521/507 , C12Q2531/119
摘要： The present invention features novel, diverse, hybrid and engineered recombinase enzymes, and the utility of such proteins with associated recombination factors for carrying out DNA amplification assays. The present invention also features different recombinase ‘systems’ having distinct biochemical activities in DNA amplification assays, and differing requirements for loading factors, single-stranded DNA binding proteins (SSBs), and the quantity of crowding agent employed.
摘要翻译： 本发明的特征在于新颖，多样，杂交和工程重组酶，以及具有相关重组因子的这种蛋白质用于进行DNA扩增测定。 本发明还具有在DNA扩增测定中具有不同生物化学活性的不同重组酶“系统”，对负载因子，单链DNA结合蛋白（SSB）以及所用的拥挤剂的量的不同要求。
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公开(公告)号：US09157127B2
公开(公告)日：2015-10-13
申请号：US14183113
申请日：2014-02-18
CPC分类号： C12Q1/6851 , C12Q1/6844 , C12Q1/689 , C12Q2600/158 , G01N21/6428 , G01N21/6456 , G01N2021/6432 , Y02A50/53 , C12Q2521/507 , C12Q2522/101 , C12Q2527/137 , C12Q2565/629 , C12Q2563/155 , C12Q2563/159
摘要： A process includes providing a mixture that includes a recombinase, a single-strand binding protein, and one or more oligonucleotides; and detecting particles in the reaction mixture.
摘要翻译： 一种方法包括提供包含重组酶，单链结合蛋白和一种或多种寡核苷酸的混合物; 并检测反应混合物中的颗粒。
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