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公开(公告)号：US20050186588A1
公开(公告)日：2005-08-25
申请号：US10967711
申请日：2004-10-18
申请人： Victor Lyamichev , Andrew Lukowiak , Nancy Jarvis , David Kurensky , Hatim Allawi , Scott Law , Chad Gerber , Robert Roeven , Jeff Hall
发明人： Victor Lyamichev , Andrew Lukowiak , Nancy Jarvis , David Kurensky , Hatim Allawi , Scott Law , Chad Gerber , Robert Roeven , Jeff Hall
IPC分类号： C12P19/34 , C12Q20060101 , C12Q1/68
CPC分类号： C12Q1/6846 , C12Q2527/119 , C12Q2527/125 , C12Q2561/109
摘要： The present invention provides methods and routines for developing and optimizing nucleic acid detection assays for use in basic research, clinical research, and for the development of clinical detection assays. In particular, the present invention provides methods for designing oligonucleotide primers to be used in multiplex amplification reactions. The present invention also provides methods to optimize multiplex amplification reactions. The present invention also provides methods for combined target and signal generation assays.
摘要翻译： 本发明提供用于开发和优化用于基础研究，临床研究和用于开发临床检测测定的核酸检测测定的方法和程序。 特别地，本发明提供了用于多重扩增反应中使用的寡核苷酸引物的设计方法。 本发明还提供优化多重扩增反应的方法。 本发明还提供了用于组合的靶和信号产生测定的方法。
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公开(公告)号：US20110104682A1
公开(公告)日：2011-05-05
申请号：US12898490
申请日：2010-10-05
申请人： Hatim Allawi , Brad T. Argue , Christian Tor Bartholomay , LuAnne Chehak , Michelle L. Curtis , Peggy S. Eis , Jeff G. Hall , Hon S. Ip , Lin Ji , Michael Kaiser , Robert W. Kwiatkowski, JR. , Andrew A. Lukowiak , Victor Lyamichev , Natalie E. Lyamicheva , WuPo Ma , Bruce P. Neri , Sarah M. Olson , Marilyn C. Olson-Munoz , James J. Schaefer , Zbigniev Skrzypczynski , Tsetska Y. Takova , Lisa C. Thompson , Kevin L. Vedvik
发明人： Hatim Allawi , Brad T. Argue , Christian Tor Bartholomay , LuAnne Chehak , Michelle L. Curtis , Peggy S. Eis , Jeff G. Hall , Hon S. Ip , Lin Ji , Michael Kaiser , Robert W. Kwiatkowski, JR. , Andrew A. Lukowiak , Victor Lyamichev , Natalie E. Lyamicheva , WuPo Ma , Bruce P. Neri , Sarah M. Olson , Marilyn C. Olson-Munoz , James J. Schaefer , Zbigniev Skrzypczynski , Tsetska Y. Takova , Lisa C. Thompson , Kevin L. Vedvik
CPC分类号： C12Q1/6823 , C12N9/1252 , C12N9/22 , C12Q1/683 , Y10T436/143333 , C12Q2561/109
摘要： The present invention provides novel cleavage agents and polymerases for the cleavage and modification of nucleic acid. The cleavage agents and polymerases find use, for example, for the detection and characterization of nucleic acid sequences and variations in nucleic acid sequences. In some embodiments, the 5′ nuclease activity of a variety of enzymes is used to cleave a target-dependent cleavage structure, thereby indicating the presence of specific nucleic acid sequences or specific variations thereof.
摘要翻译： 本发明提供了用于切割和修饰核酸的新型切割剂和聚合酶。 裂解剂和聚合酶可用于例如核酸序列的检测和表征以及核酸序列的变化。 在一些实施方案中，使用各种酶的5'核酸酶活性来切割靶依赖性切割结构，从而指示特异性核酸序列的存在或其特定变体。
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公开(公告)号：US20060199202A1
公开(公告)日：2006-09-07
申请号：US11349872
申请日：2006-02-08
申请人： Victor Lyamichev , Hatim Allawi
发明人： Victor Lyamichev , Hatim Allawi
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6827 , C12Q2561/109 , C12Q2545/114
摘要： The present invention provides compositions and methods for the detection and characterization of allelic expression imbalance from a heterozygous gene locus. More particularly, the present invention provides compositions, kits, and methods for the determination of allelic expression imbalance by the comparison of expression levels from each of two alleles of a given gene locus through the use of an invasive cleavage structure assay (e.g. the INVADER assay).
摘要翻译： 本发明提供了用于检测和表征杂合基因座的等位基因表达不平衡的组合物和方法。 更具体地，本发明通过使用侵入性切割结构测定法（例如INVADER测定法）来比较来自给定基因位点的两个等位基因中的每一个的表达水平来确定等位基因表达失衡的组合物，试剂盒和方法 ）。
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公开(公告)号：US07060436B2
公开(公告)日：2006-06-13
申请号：US09882945
申请日：2001-06-15
申请人： Victor Lyamichev , Hatim Allawi , Fang Dong , Bruce P. Neri , Tatiani I. Vener
发明人： Victor Lyamichev , Hatim Allawi , Fang Dong , Bruce P. Neri , Tatiani I. Vener
CPC分类号： C12Q1/6827 , C12Q1/6811 , C12Q1/686 , C12Q1/703 , C12Q2535/125 , C12Q2525/301
摘要： The present invention relates to methods and compositions for analyzing nucleic acids, and in particular, methods and compositions for detection and characterization of nucleic acid sequences and sequence changes. The present invention also provides methods and compositions for identifying oligonucleotides with desired hybridization properties to nucleic acid targets containing secondary structure.
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公开(公告)号：US08361720B2
公开(公告)日：2013-01-29
申请号：US12946737
申请日：2010-11-15
CPC分类号： C12Q1/6827 , C12Q1/6818 , C12Q1/686 , C12Q1/6886 , C12Q2600/156 , C12Q2600/158 , C12Q2525/301 , C12Q2527/101 , C12Q2561/109 , C12Q2565/1015
摘要： A cleavage-based real-time PCR assay method is provided. In general terms, the assay method includes subjecting a reaction mixture comprising a) PCR reagents for amplifying a nucleic acid target, and b) flap cleavage reagents for performing a flap cleavage assay on the amplified nucleic acid target to two sets of thermocycling conditions. No additional reagents are added to the reaction between said first and second sets of cycles and, in each cycle of the second set of cycles, cleavage of a flap probe is measured.
摘要翻译： 提供基于切割的实时PCR测定方法。 一般而言，测定方法包括对包含a）PCR试剂的反应混合物进行核酸靶扩增，以及b）用于在扩增的核酸靶上进行皮瓣裂解测定的折叠裂解试剂至两组热循环条件。 在所述第一组和第二组循环之间没有向所述反应中加入另外的试剂，并且在第二组循环的每个循环中，测量瓣探针的切割。
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公开(公告)号：US08916344B2
公开(公告)日：2014-12-23
申请号：US12946745
申请日：2010-11-15
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6886 , C12Q1/6827 , C12Q2600/154 , C12Q2521/301 , C12Q2523/125 , C12Q2561/109
摘要： A method for detecting a methylated genomic locus is provided. In certain embodiments, the method comprises: a) treating a nucleic acid sample that contains both unmethylated and methylated copies of a genomic locus with an agent that modifies cytosine to uracil to produce a treated nucleic acid; b) amplifying a product from the treated nucleic acid using a first primer and a second primer, wherein the first primer hybridizes to a site in the locus that contain methylcytosines and the amplifying preferentially amplifies the methylated copies of the genomic locus, to produce an amplified sample; and c) detecting the presence of amplified methylated copies of the genomic locus in the amplified sample using a flap assay that employs an invasive oligonucleotide having a 3′ terminal G or C nucleotide that corresponds to a site of methylation in the genomic locus.
摘要翻译： 提供了一种检测甲基化基因组座位的方法。 在某些实施方案中，所述方法包括：a）将含有基因组座位的非甲基化和甲基化拷贝的核酸样品用修饰胞嘧啶至尿嘧啶的试剂处理以产生经处理的核酸; b）使用第一引物和第二引物扩增来自经处理的核酸的产物，其中第一引物与含有甲基胞嘧啶的位点的位点杂交，并且扩增优先扩增基因组基因座的甲基化拷贝，以产生扩增 样品; 和c）使用采用具有对应于基因组基因座中的甲基化位点的3'末端G或C核苷酸的侵入性寡核苷酸的皮瓣测定来检测扩增样品中扩增的基因组座位的扩增的甲基化拷贝的存在。
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公开(公告)号：US20130231256A1
公开(公告)日：2013-09-05
申请号：US13594674
申请日：2012-08-24
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6886 , C12Q1/6858 , C12Q2600/156 , C12Q2600/16 , C12Q2525/161 , C12Q2561/109 , C12Q2565/1015
摘要： Provided herein is reagent mixture comprising multiplexed amplification reagents and flap assay reagents for detecting, in a single reaction, mutant copies of the KRAS gene that contain any of the 34A, 34C, 34T, 35A, 35C, 35T or 38A point mutations. Methods that employ the reagent mix and kits for performing the same are also provided.
摘要翻译： 本文提供的试剂混合物包含多重扩增试剂和皮瓣测定试剂，用于在单次反应中检测含有34A，34C，34T，35A，35C，35T或38A点突变中任何一种的KRAS基因的突变体拷贝。 还提供了使用试剂混合物和试剂盒进行其的方法。
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公开(公告)号：US20120122106A1
公开(公告)日：2012-05-17
申请号：US12946752
申请日：2010-11-15
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6886 , C12Q1/6858 , C12Q1/6881 , C12Q2600/156 , C12Q2600/158 , C12Q2525/301 , C12Q2561/109 , C12Q2565/1015
摘要： A method of sample analysis is provided. In certain embodiments, the method involves: a) amplifying a product from a sample that comprises both wild type copies of a genomic locus and mutant copies of the genomic locus that have a point mutation relative to said wild type copies of the genomic locus, to produce an amplified sample, where: i. the amplifying is done using a first primer and a second primer; and ii. the first primer comprises a 3′ terminal nucleotide that base pairs with the point mutation and also comprises a nucleotide sequence that is fully complementary to a sequence in the locus with the exception of a single base mismatch within 6 bases of the 3′ terminal nucleotide; and b) detecting the presence of said product in said amplified sample using a flap assay that employs an invasive oligonucleotide. A kit for performing the method is also provided.
摘要翻译： 提供了样品分析的方法。 在某些实施方案中，该方法包括：a）从包含基因组基因组的野生型拷贝和相对于所述基因组基因座的所述野生型拷贝的点突变的基因组基因座的突变拷贝的样品扩增产物， 产生扩增样品，其中：i。 使用第一引物和第二引物进行扩增; 和ii。 第一引物包含与点突变碱基对的3'末端核苷酸，并且还包含与3'末端核苷酸的6个碱基内的单碱基错配除外的基因座中的序列完全互补的核苷酸序列; 和b）使用采用侵入性寡核苷酸的皮瓣测定来检测所述扩增样品中所述产物的存在。 还提供了用于执行该方法的套件。
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公开(公告)号：US20120122088A1
公开(公告)日：2012-05-17
申请号：US12946745
申请日：2010-11-15
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6886 , C12Q1/6827 , C12Q2600/154 , C12Q2521/301 , C12Q2523/125 , C12Q2561/109
摘要： A method for detecting a methylated genomic locus is provided. In certain embodiments, the method comprises: a) treating a nucleic acid sample that contains both unmethylated and methylated copies of a genomic locus with an agent that modifies cytosine to uracil to produce a treated nucleic acid; b) amplifying a product from the treated nucleic acid using a first primer and a second primer, wherein the first primer hybridizes to a site in the locus that contain methylcytosines and the amplifying preferentially amplifies the methylated copies of the genomic locus, to produce an amplified sample; and c) detecting the presence of amplified methylated copies of the genomic locus in the amplified sample using a flap assay that employs an invasive oligonucleotide having a 3′ terminal G or C nucleotide that corresponds to a site of methylation in the genomic locus.
摘要翻译： 提供了一种检测甲基化基因组座位的方法。 在某些实施方案中，所述方法包括：a）将含有基因组座位的非甲基化和甲基化拷贝的核酸样品用修饰胞嘧啶至尿嘧啶的试剂处理以产生经处理的核酸; b）使用第一引物和第二引物扩增来自经处理的核酸的产物，其中第一引物与含有甲基胞嘧啶的位点的位点杂交，并且扩增优先扩增基因组基因座的甲基化拷贝，以产生扩增的 样品; 和c）使用采用具有对应于基因组基因座中的甲基化位点的3'末端G或C核苷酸的侵入性寡核苷酸的皮瓣测定来检测扩增样品中扩增的基因组座位的扩增的甲基化拷贝的存在。
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公开(公告)号：US09127318B2
公开(公告)日：2015-09-08
申请号：US13594674
申请日：2012-08-24
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6886 , C12Q1/6858 , C12Q2600/156 , C12Q2600/16 , C12Q2525/161 , C12Q2561/109 , C12Q2565/1015
摘要： Provided herein is reagent mixture comprising multiplexed amplification reagents and flap assay reagents for detecting, in a single reaction, mutant copies of the KRAS gene that contain any of the 34A, 34C, 34T, 35A, 35C, 35T or 38A point mutations. Methods that employ the reagent mix and kits for performing the same are also provided.
摘要翻译： 本文提供的试剂混合物包含多重扩增试剂和皮瓣测定试剂，用于在单次反应中检测含有34A，34C，34T，35A，35C，35T或38A点突变中任何一种的KRAS基因的突变体拷贝。 还提供了使用试剂混合物和试剂盒进行其的方法。
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