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1. 发明授权

US07132239B2 Determination of a genotype of an amplification product at multiple allelic sites 有权
标题翻译：确定多个等位基因位点扩增产物的基因型
公开(公告)号：US07132239B2
公开(公告)日：2006-11-07
申请号：US10455150
申请日：2003-06-04
申请人： Kenneth J. Livak , Federico Goodsaid
发明人： Kenneth J. Livak , Federico Goodsaid
IPC分类号： C12Q1/68 , C12P19/34 , C07H21/02
CPC分类号： C12Q1/6858 , C12Q1/6818 , C12Q2561/101 , C12Q2537/143 , C12Q2563/107 , C12Q2531/119
摘要： A method is provided for genotyping a target sequence at at least two allelic sites by a 5′ nuclease amplification reaction. In one embodiment, the method includes performing a nucleic acid amplification on a target sequence having at least two different allelic sites using a nucleic acid polymerase having 5′→3′ nuclease activity and a primer capable of hybridizing to the target sequence in the presence of two or more sets of allelic oligonucleotide probes wherein: each set of allelic oligonucleotide probes is for detecting a different allelic site of the target sequence, each set of allelic oligonucleotide probes includes two or more probes which are complementary to different allelic variants at the allelic site being detected by the set of probes, the allelic site being 5′ relative to a sequence to which the primer hybridizes to the target sequence, and at least all but one of the allelic oligonucleotide probes include a different fluorescer than the other probes and a quencher positioned on the probe to quench the fluorescence of the fluorescer; detecting a fluorescence spectrum of the amplification;calculating a fluorescence contribution of each fluorescer to the fluorescence spectrum; anddetermining a presence or absence of the different allelic variants at the two or more different allelic sites based on the fluorescence contribution of each fluorescer to the combined fluorescence spectrum.
摘要翻译：提供了一种通过5'核酸酶扩增反应在至少两个等位基因位点对靶序列进行基因分型的方法。在一个实施方案中，该方法包括使用具有5'→3'核酸酶活性的核酸聚合酶和能够在目标序列中与靶序列杂交的引物对具有至少两个不同等位基因位点的靶序列进行核酸扩增两组或更多组等位基因寡核苷酸探针，其中：每组等位基因寡核苷酸探针用于检测靶序列的不同等位基因位点，每组等位基因寡核苷酸探针包括与等位基因上的不同等位基因变体互补的两个或多个探针通过该组探针检测位点，所述等位基因位点相对于引物与靶序列杂交的序列为5'，并且至少所有等位基因寡核苷酸探针中的所有等位基因寡核苷酸探针包括与其它探针不同的荧光剂，和猝灭剂位于探针上以淬灭荧光剂的荧光; 检测扩增的荧光光谱; 计算每个荧光剂对荧光光谱的荧光贡献; 并且基于每个荧光剂对组合的荧光光谱的荧光贡献，确定两个或更多个不同等位基因位点处的不同等位基因变体的存在或不存在。

2. 发明授权

US06884583B2 Determination of a genotype of an amplification product at multiple allelic sites 有权
标题翻译：确定多个等位基因位点扩增产物的基因型
公开(公告)号：US06884583B2
公开(公告)日：2005-04-26
申请号：US10104774
申请日：2002-03-21
申请人： Kenneth J. Livak , Federico Goodsaid
发明人： Kenneth J. Livak , Federico Goodsaid
IPC分类号： G01N21/64 , C12M1/00 , C12Q1/68 , G01N33/53 , G01N33/566 , C07H21/02 , C12P19/34
CPC分类号： C12Q1/6858 , C12Q1/6818 , C12Q2561/101 , C12Q2537/143 , C12Q2563/107 , C12Q2531/119
摘要： A method is provided for genotyping a target sequence at at least two allelic sites by a 5′ nuclease amplification reaction. In one embodiment, the method includes performing a nucleic acid amplification on a target sequence having at least two different allelic sites using a nucleic acid polymerase having 5′→3′ nuclease activity and a primer capable of hybridizing to the target sequence in the presence of two or more sets of allelic oligonucleotide probes wherein: each set of allelic oligonucleotide probes is for detecting a different allelic site of the target sequence, each set of allelic oligonucleotide probes includes two or more probes which are complementary to different allelic variants at the allelic site being detected by the set of probes, the allelic site being 5′ relative to a sequence to which the primer hybridizes to the target sequence, and at least all but one of the allelic oligonucleotide probes include a different fluorescer than the other probes and a quencher postioned on the probe to quench the fluorescence of the fluorescer; detecting a fluorescence spectrum of the amplification; calculating a fluorescence contribution of each fluorescer to the fluorescence spectrum; and determining a presence or absence of the different allelic variants at the two or more different allelic sites based on the fluorescence contribution of each fluorescer to the combined fluorescence spectrum.
摘要翻译：提供了一种通过5'核酸酶扩增反应在至少两个等位基因位点对靶序列进行基因分型的方法。在一个实施方案中，该方法包括使用具有5'→3'核酸酶活性的核酸聚合酶和能够在目标序列中与靶序列杂交的引物对具有至少两个不同等位基因位点的靶序列进行核酸扩增两组或更多组等位基因寡核苷酸探针，其中：每组等位基因寡核苷酸探针用于检测靶序列的不同等位基因位点，每组等位基因寡核苷酸探针包括与等位基因上的不同等位基因变体互补的两个或多个探针通过该组探针检测位点，所述等位基因位点相对于引物与靶序列杂交的序列为5'，并且至少所有等位基因寡核苷酸探针中的所有等位基因寡核苷酸探针包括与其它探针不同的荧光剂，和探针上的猝灭剂用于淬灭荧光剂的荧光; 检测扩增的荧光光谱; 计算每个荧光剂对荧光光谱的荧光贡献; 并且基于每个荧光剂对组合的荧光光谱的荧光贡献，确定两个或更多个不同等位基因位点处的不同等位基因变体的存在或不存在。

3. 发明申请

US20050145496A1 Thermal reaction device and method for using the same 审中-公开
公开(公告)号：US20050145496A1
公开(公告)日：2005-07-07
申请号：US10876046
申请日：2004-06-23
申请人： Federico Goodsaid , Marc Unger , Jiang Huang , Emerson Quan
发明人： Federico Goodsaid , Marc Unger , Jiang Huang , Emerson Quan
IPC分类号： B01L3/00 , G01N27/453
CPC分类号： B01L3/00 , B01L3/0248 , B01L3/5025 , B01L3/5027 , B01L3/502707 , B01L3/502715 , B01L3/50273 , B01L3/502738 , B01L7/52 , B01L2200/0642 , B01L2200/142 , B01L2200/147 , B01L2300/0636 , B01L2300/0816 , B01L2300/0819 , B01L2300/0861 , B01L2300/0864 , B01L2300/0867 , B01L2300/123 , B01L2300/1805 , B01L2400/0481 , B01L2400/0487 , B01L2400/0638 , B01L2400/0655 , C12Q1/686 , G01N27/453
摘要： An M×N matrix microfluidic device for performing a matrix of reactions, the device having a plurality of reaction cells in communication with one of either a sample inlet or a reagent inlet through a via formed within an elastomeric block of the device. Methods provided include a method for forming vias in parallel in an elastomeric layer of an elastomeric block of a microfluidic device, the method comprising using patterned photoresist masks and etching reagents to etch away regions or portions of an elastomeric layer of the elastomeric block.

4. 发明授权

US5962233A Determination of a genotype of an amplification product at multiple allelic sites 失效
公开(公告)号：US5962233A
公开(公告)日：1999-10-05
申请号：US18595
申请日：1998-02-04
申请人： Kenneth J. Livak , Federico Goodsaid
发明人： Kenneth J. Livak , Federico Goodsaid
摘要： A method is provided for genotyping a target sequence at at least two allelic sites by a 5' nuclease amplification reaction. In one embodiment, the method includes performing a nucleic acid amplification on a target sequence having at least two different allelic sites using a nucleic acid polymerase having 5'.fwdarw.3' nuclease activity and a primer capable of hybridizing to the target sequence in the presence of two or more sets of allelic oligonucleotide probes wherein:each set of allelic oligonucleotide probes is for detecting a different allelic site of the target sequence,each set of allelic oligonucleotide probes includes two or more probes which are complementary to different allelic variants at the allelic site being detected by the set of probes, the allelic site being 5' relative to a sequence to which the primer hybridizes to the target sequence, andat least all but one of the allelic oligonucleotide probes include a different fluorescer than the other probes and a quencher positioned on the probe to quench the fluorescence of the fluorescer;detecting a fluorescence spectrum of the amplification;calculating a fluorescence contribution of each fluorescer to the fluorescence spectrum; anddetermining a presence or absence of the different allelic variants at the two or more different allelic sites based on the fluorescence contribution of each fluorescer to the combined fluorescence spectrum.

5. 发明申请

US20120053069A1 Determination of a Genotype of an Amplification Product at Multiple Allelic Sites 审中-公开
标题翻译：确定多个等位基因位点扩增产物的基因型
公开(公告)号：US20120053069A1
公开(公告)日：2012-03-01
申请号：US12618686
申请日：2009-11-13
申请人： Kenneth J. LIVAK , Federico Goodsaid
发明人： Kenneth J. LIVAK , Federico Goodsaid
IPC分类号： C40B30/04 , C40B40/06 , C12Q1/68
CPC分类号： C12Q1/6858 , C12Q1/6818 , C12Q2561/101 , C12Q2537/143 , C12Q2563/107 , C12Q2531/119
摘要： A method is provided for genotyping a target sequence at at least two allelic sites by a 5′ nuclease amplification reaction. In one embodiment, the method includes performing a nucleic acid amplification on a target sequence having at least two different allelic sites using a nucleic acid polymerase having 5′-3′ nuclease activity and a primer capable of hybridizing to the target sequence in the presence of two or more sets of allelic oligonucleotide probes wherein at least all but one of the allelic oligonucleotide probes include a different fluorescer than the other probes and a quencher positioned on the probe to quench the fluorescence of the fluorescer; detecting a fluorescence spectrum of the amplification; calculating a fluorescence contribution of each fluorescer to the fluorescence spectrum; and determining a presence or absence of the different allelic variants based on the fluorescence contribution of each fluorescer to the combined fluorescence spectrum.
摘要翻译：提供了一种通过5'核酸酶扩增反应在至少两个等位基因位点对靶序列进行基因分型的方法。在一个实施方案中，该方法包括使用具有5'-3'核酸酶活性的核酸聚合酶和能够与靶序列杂交的引物在具有至少两个不同等位基因位点的靶序列上进行核酸扩增两组或多组等位基因寡核苷酸探针，其中等位基因寡核苷酸探针中的至少全部除一个之外包括与其它探针不同的荧光剂，以及位于探针上的猝灭剂以淬灭荧光剂的荧光; 检测扩增的荧光光谱; 计算每个荧光剂对荧光光谱的荧光贡献; 并且基于每个荧光剂对组合的荧光光谱的荧光贡献来确定不同等位基因变体的存在或不存在。

6. 发明授权

US6154707A Computer logic for fluorescence genotyping at multiple allelic sites 有权
标题翻译：用于多个等位基因位点荧光基因分型的计算机逻辑
公开(公告)号：US6154707A
公开(公告)日：2000-11-28
申请号：US324709
申请日：1999-06-03
申请人： Kenneth J. Livak , Federico Goodsaid
发明人： Kenneth J. Livak , Federico Goodsaid
IPC分类号： G01N21/64 , C12M1/00 , C12Q1/68 , G01N33/53 , G01N33/566 , G06G7/58
CPC分类号： C12Q1/6858 , C12Q1/6818
摘要： A method is provided for genotyping a target sequence at at least two allelic sites by a 5' nuclease amplification reaction. In one embodiment, the method includes performing a nucleic acid amplification on a target sequence having at least two different allelic sites using a nucleic acid polymerase having 5'.fwdarw.3' nuclease activity and a primer capable of hybridizing to the target sequence in the presence of two or more sets of allelic oligonucleotide probes wherein:each set of allelic oligonucleotide probes is for detecting a different allelic site of the target sequence,each set of allelic oligonucleotide probes includes two or more probes which are complementary to different allelic variants at the allelic site being detected by the set of probes, the allelic site being 5' relative to a sequence to which the primer hybridizes to the target sequence, andat least all but one of the allelic oligonucleotide probes include a different fluorescer than the other probes and a quencher positioned on the probe to quench the fluorescence of the fluorescer;detecting a fluorescence spectrum of the amplification;calculating a fluorescence contribution of each fluorescer to the fluorescence spectrum; anddetermining a presence or absence of the different allelic variants at the two or more different allelic sites based on the fluorescence contribution of each fluorescer to the combined fluorescence spectrum.
摘要翻译：提供了一种通过5'核酸酶扩增反应在至少两个等位基因位点对靶序列进行基因分型的方法。在一个实施方案中，该方法包括使用具有5'→3'核酸酶活性的核酸聚合酶和能够在目标序列中与靶序列杂交的引物对具有至少两个不同等位基因位点的靶序列进行核酸扩增两组或更多组等位基因寡核苷酸探针，其中：每组等位基因寡核苷酸探针用于检测靶序列的不同等位基因位点，每组等位基因寡核苷酸探针包括与等位基因上的不同等位基因变体互补的两个或多个探针通过该组探针检测位点，所述等位基因位点相对于引物与靶序列杂交的序列为5'，并且至少所有等位基因寡核苷酸探针中的所有等位基因寡核苷酸探针包括与其它探针不同的荧光剂，和猝灭剂位于探针上以淬灭荧光剂的荧光; 检测扩增的荧光光谱; 计算每个荧光剂对荧光光谱的荧光贡献; 并且基于每个荧光剂对组合的荧光光谱的荧光贡献，确定两个或更多个不同等位基因位点处的不同等位基因变体的存在或不存在。

7. 发明授权

US07867454B2 Thermal reaction device and method for using the same 有权
标题翻译：热反应装置及其使用方法
公开(公告)号：US07867454B2
公开(公告)日：2011-01-11
申请号：US11929370
申请日：2007-10-30
申请人： Federico Goodsaid , Marc Unger , Jiang Huang , Emerson Quan
发明人： Federico Goodsaid , Marc Unger , Jiang Huang , Emerson Quan
IPC分类号： B01L3/00
CPC分类号： B01L3/00 , B01L3/0248 , B01L3/5025 , B01L3/5027 , B01L3/502707 , B01L3/502715 , B01L3/50273 , B01L3/502738 , B01L7/52 , B01L2200/0642 , B01L2200/142 , B01L2200/147 , B01L2300/0636 , B01L2300/0816 , B01L2300/0819 , B01L2300/0861 , B01L2300/0864 , B01L2300/0867 , B01L2300/123 , B01L2300/1805 , B01L2400/0481 , B01L2400/0487 , B01L2400/0638 , B01L2400/0655 , C12Q1/686 , G01N27/453
摘要： An M.times.N matrix microfluidic device for performing a matrix of reactions, the device having a plurality of reaction cells in communication with one of either a sample inlet or a reagent inlet through a via formed within an elastomeric block of the device. Methods provided include a method for forming vias in parallel in an elastomeric layer of an elastomeric block of a microfluidic device, the method comprising using patterned photoresist masks and etching reagents to etch away regions or portions of an elastomeric layer of the elastomeric block.
摘要翻译：一种用于执行反应矩阵的M.times.N矩阵微流体装置，该装置具有多个反应池，其通过形成在装置的弹性体块内的通孔与样品入口或试剂入口之一连通。提供的方法包括在微流体装置的弹性体块的弹性体层中平行形成通孔的方法，该方法包括使用图案化的光致抗蚀剂掩模和蚀刻试剂来蚀刻掉弹性体块的弹性体层的区域或部分。

8. 发明申请

US20080292504A1 Thermal Reaction Device and Method for Using the Same 有权
标题翻译：热反应装置及其使用方法
公开(公告)号：US20080292504A1
公开(公告)日：2008-11-27
申请号：US11929370
申请日：2007-10-30
申请人： Federico Goodsaid , Marc Unger , Jiang Huang , Emerson Quan
发明人： Federico Goodsaid , Marc Unger , Jiang Huang , Emerson Quan
IPC分类号： G01N21/64 , B01J19/00
CPC分类号： B01L3/00 , B01L3/0248 , B01L3/5025 , B01L3/5027 , B01L3/502707 , B01L3/502715 , B01L3/50273 , B01L3/502738 , B01L7/52 , B01L2200/0642 , B01L2200/142 , B01L2200/147 , B01L2300/0636 , B01L2300/0816 , B01L2300/0819 , B01L2300/0861 , B01L2300/0864 , B01L2300/0867 , B01L2300/123 , B01L2300/1805 , B01L2400/0481 , B01L2400/0487 , B01L2400/0638 , B01L2400/0655 , C12Q1/686 , G01N27/453
摘要： An M.times.N matrix microfluidic device for performing a matrix of reactions, the device having a plurality of reaction cells in communication with one of either a sample inlet or a reagent inlet through a via formed within an elastomeric block of the device. Methods provided include a method for forming vias in parallel in an elastomeric layer of an elastomeric block of a microfluidic device, the method comprising using patterned photoresist masks and etching reagents to etch away regions or portions of an elastomeric layer of the elastomeric block.
摘要翻译：一种用于执行反应矩阵的M.times.N矩阵微流体装置，该装置具有多个反应池，其通过形成在装置的弹性体块内的通孔与样品入口或试剂入口之一连通。提供的方法包括在微流体装置的弹性体块的弹性体层中平行形成通孔的方法，该方法包括使用图案化的光致抗蚀剂掩模和蚀刻试剂来蚀刻掉弹性体块的弹性体层的区域或部分。

9. 发明申请

US20080187913A1 Determination of genotype of an amplification product at multiple allelic sites 审中-公开
标题翻译：确定多个等位基因位点扩增产物的基因型
公开(公告)号：US20080187913A1
公开(公告)日：2008-08-07
申请号：US11592716
申请日：2006-11-03
申请人： Kenneth J. Livak , Federico Goodsaid
发明人： Kenneth J. Livak , Federico Goodsaid
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6858 , C12Q1/6818 , C12Q2561/101 , C12Q2537/143 , C12Q2563/107 , C12Q2531/119
摘要： A method is provided for genotyping a target sequence at least two allelic sites by a 5′ nuclease amplification reaction. In one embodiment, the method includes performing a nucleic acid amplification on a target sequence having at least two different allelic sites using a nucleic acid polymerase having 5′-3′ nuclease activity and a primer capable of hybridizing to the target sequence in the presence of two or more sets of allelic oligonucleotide probes wherein: each set of allelic oligonucleotide probes is for detecting a different allelic site of the target sequence, each set of allelic oligonucleotide probes includes two or more probes which are complementary to different allelic variants at the allelic site being detected by the set of probes, the allelic site being 5′ relative to a sequence to which the primer hybridizes to the target sequence, and at least all but one of the allelic oligonucleotide probes include a different fluorescer than the other probes and a quencher positioned on the probe to quench the fluorescence of the fluorescer; detecting a fluorescence spectrum of the amplification; calculating a fluorescence contribution of each fluorescer to the fluorescence spectrum; and determining a presence or absence of the different allelic variants at the two or more different allelic sites based on the fluorescence contribution of each fluorescer to the combined fluorescence spectrum.
摘要翻译：提供了一种通过5'核酸酶扩增反应将靶序列至少两个等位基因位点进行基因分型的方法。在一个实施方案中，该方法包括使用具有5'-3'核酸酶活性的核酸聚合酶和能够与靶序列杂交的引物在具有至少两个不同等位基因位点的靶序列上进行核酸扩增两组或更多组等位基因寡核苷酸探针，其中：每组等位基因寡核苷酸探针用于检测靶序列的不同等位基因位点，每组等位基因寡核苷酸探针包括与等位基因位点上的不同等位基因变体互补的两个或多个探针被该组探针检测到，等位基因位点相对于引物与靶序列杂交的序列为5'，并且至少所有等位基因寡核苷酸探针中的所有等位基因寡核苷酸探针包括与其它探针不同的荧光剂和猝灭剂位于探针上以淬灭荧光剂的荧光; 检测扩增的荧光光谱; 计算每个荧光剂对荧光光谱的荧光贡献; 并且基于每个荧光剂对组合的荧光光谱的荧光贡献，确定两个或更多个不同等位基因位点处的不同等位基因变体的存在或不存在。

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