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公开(公告)号:US09000146B2
公开(公告)日:2015-04-07
申请号:US13470251
申请日:2012-05-11
申请人: Janelle J. Bruinsma , Michael J. Domanico , Graham P. Lidgard , Hongzhi Zou , William G. Weisburg , Hemanth D. Shenoi , James P. Light, II
发明人: Janelle J. Bruinsma , Michael J. Domanico , Graham P. Lidgard , Hongzhi Zou , William G. Weisburg , Hemanth D. Shenoi , James P. Light, II
CPC分类号: C12Q1/6806 , B01D33/15 , B01D33/155 , B01L3/00 , B01L3/5021 , B03C1/30 , B04B3/00 , C12N15/1006 , C12N15/1013 , C12N15/1017 , C12Q1/6886 , C12Q2600/158 , C12Q2600/16 , Y10T436/143333
摘要: Provided herein is technology relating to isolating nucleic acids. In particular, the technology relates to methods and kits for extracting multiple DNA targets from a human stool sample with, for example, methods employing spin filtration techniques.
摘要翻译: 本文提供了与分离核酸有关的技术。 特别地,该技术涉及用于从例如使用旋转过滤技术的方法从人粪便样品中提取多个DNA靶标的方法和试剂盒。
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公开(公告)号:US20100297658A1
公开(公告)日:2010-11-25
申请号:US12851872
申请日:2010-08-06
申请人: David A. Ahlquist , Hongzhi Zou
发明人: David A. Ahlquist , Hongzhi Zou
CPC分类号: C12Q1/6806 , C12N15/1003 , C12Q1/6886 , C12Q2600/154
摘要: This document includes methods and materials for enriching and detecting cancer markers. For example, this document includes methods and materials for enriching methylated mammalian nucleic acid from stool samples.
摘要翻译: 本文件包括丰富和检测癌症标志物的方法和材料。 例如,本文件包括从粪便样品中富集甲基化的哺乳动物核酸的方法和材料。
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公开(公告)号:US20100121046A1
公开(公告)日:2010-05-13
申请号:US12555672
申请日:2009-09-08
申请人: David A. Ahlquist , Jonathan J. Harrington , Hongzhi Zou , Patrick S. Quint , William R. Taylor
发明人: David A. Ahlquist , Jonathan J. Harrington , Hongzhi Zou , Patrick S. Quint , William R. Taylor
CPC分类号: B01L3/00 , A61B10/0038 , A61B10/0096 , B01L3/5029 , C12Q1/6806 , C12Q2565/507 , C12Q2563/143 , C12Q2527/125
摘要: This document provides methods and materials involved in collecting and processing complex macromolecular mixtures (e.g., stool samples). For example, stool collection devices, buffers for stabilizing nucleic acid and polypeptides present in stool, and kits for using sequence-specific capture probes (e.g., nucleic acid sequences designed to hybridize with particular target nucleic acids) to capture target nucleic acids directly from complex macromolecular mixtures (e.g., stool samples) without the need to perform prior steps to enrich, isolate, or purify the nucleic acid component are provided.
摘要翻译: 本文件提供了收集和处理复杂大分子混合物(例如粪便样品)中涉及的方法和材料。 例如,粪便收集装置,用于稳定存在于粪便中的核酸和多肽的缓冲液以及使用序列特异性捕获探针(例如,设计为与特定靶核酸杂交的核酸序列)直接从复合物捕获靶核酸的试剂盒 提供大分子混合物(例如,粪便样品),而不需要执行先前的富集,分离或纯化核酸组分的步骤。
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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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公开(公告)号:US08715937B2
公开(公告)日:2014-05-06
申请号:US12946752
申请日:2010-11-15
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 the 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 the product in the 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,所述方法包括:a)从包含基因组基因组的野生型拷贝和相对于基因组基因座的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法检测扩增样品中产物的存在。 还提供了用于执行该方法的套件。
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公开(公告)号:US20120122105A1
公开(公告)日:2012-05-17
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切割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:US20110183328A1
公开(公告)日:2011-07-28
申请号:US12866558
申请日:2009-02-11
申请人: William R. Taylor , Jonathan J. Harrington , Patrick S. Quint , Hongzhi Zou , Harold R. Bergen, III , David I. Smith , David A. Ahlquist
发明人: William R. Taylor , Jonathan J. Harrington , Patrick S. Quint , Hongzhi Zou , Harold R. Bergen, III , David I. Smith , David A. Ahlquist
CPC分类号: C12Q1/6886 , C12Q1/37 , C12Q1/40 , C12Q2600/154 , C12Q2600/156 , C12Q2600/158 , C12Y302/01001 , C12Y304/2107 , G01N33/57407 , G01N33/57419 , G01N33/57438 , G01N2333/928 , G01N2333/966 , G01N2560/00
摘要: This document relates to methods and materials for detecting premalignant and malignant neoplasms. For example, methods and materials for determining whether or not a stool sample from a mammal contains nucleic acid markers or polypeptide markers of a neoplasm are provided.
摘要翻译: 本文件涉及检测恶变前期和恶性肿瘤的方法和材料。 例如,提供了用于确定来自哺乳动物的粪便样品是否含有肿瘤的核酸标记物或多肽标记物的方法和材料。
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公开(公告)号:US07785772B2
公开(公告)日:2010-08-31
申请号:US12018273
申请日:2008-01-23
申请人: David A. Ahlquist , Hongzhi Zou
发明人: David A. Ahlquist , Hongzhi Zou
CPC分类号: C12Q1/6806 , C12N15/1003 , C12Q1/6886 , C12Q2600/154
摘要: This document includes methods and materials for enriching and detecting cancer markers. For example, this document includes methods and materials for enriching methylated mammalian nucleic acid from stool samples.
摘要翻译: 本文件包括丰富和检测癌症标志物的方法和材料。 例如,本文件包括从粪便样品中富集甲基化的哺乳动物核酸的方法和材料。
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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:US08673555B2
公开(公告)日:2014-03-18
申请号:US12866558
申请日:2009-02-11
申请人: William R. Taylor , Jonathan J. Harrington , Patrick S. Quint , Hongzhi Zou , Harold R. Bergen, III , David I. Smith , David A. Ahlquist
发明人: William R. Taylor , Jonathan J. Harrington , Patrick S. Quint , Hongzhi Zou , Harold R. Bergen, III , David I. Smith , David A. Ahlquist
IPC分类号: C12Q1/68
CPC分类号: C12Q1/6886 , C12Q1/37 , C12Q1/40 , C12Q2600/154 , C12Q2600/156 , C12Q2600/158 , C12Y302/01001 , C12Y304/2107 , G01N33/57407 , G01N33/57419 , G01N33/57438 , G01N2333/928 , G01N2333/966 , G01N2560/00
摘要: This document relates to methods and materials for detecting premalignant and malignant neoplasms. For example, methods and materials for determining whether or not a stool sample from a mammal contains nucleic acid markers or polypeptide markers of a neoplasm are provided.
摘要翻译: 本文件涉及检测恶变前期和恶性肿瘤的方法和材料。 例如,提供了用于确定来自哺乳动物的粪便样品是否含有肿瘤的核酸标记物或多肽标记物的方法和材料。
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