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1. 发明申请

US20150176067A1 MICROARRAY SYSTEM WITH IMPROVED SEQUENCE SPECIFICITY 审中-公开
标题翻译：具有改进的序列特异性的微阵列系统
公开(公告)号：US20150176067A1
公开(公告)日：2015-06-25
申请号：US14576076
申请日：2014-12-18
申请人： Kerry B. Gunning , Mark Aaron Behlke
发明人： Kerry B. Gunning , Mark Aaron Behlke
IPC分类号： C12Q1/68 , B01J19/00
CPC分类号： C12Q1/6837 , B01J19/0046 , B01J2219/0059 , B01J2219/00608 , B01J2219/00626 , B01J2219/00722 , C12P19/34 , C12Q1/6806 , Y10T436/108331 , C12Q2525/125 , C12Q2533/101 , C12Q2521/319
摘要： The invention provides a novel array method for nucleic acid sequence detection with improved specificity which allows for detection of genetic variation, from simple SNPs (where the variation occurs at a fixed position and is of limited allelic number) to more complex sequence variation patterns (such as with multigene families or multiple genetic strains of an organism where the sequence variation between the individual members is neither fixed nor consistent). The array is comprised of short, synthetic oligonucleotide probes attached to a solid surface which are hybridized to single-stranded targets. Single stranded targets can be produced using a method that employs primers modified on the 5′ end to prohibit degradation by a 5′-exonuclease that is introduced to degrade the unprotected strand. The invention further provides for printing buffers/solutions for the immobilization of oligonucleotide probes to an array surface. The invention also provides hybridization and wash buffers and conditions to maximize hybridization specificity and signal intensity, and reduce hybridization times.
摘要翻译：本发明提供了一种用于核酸序列检测的新型阵列方法，其具有改进的特异性，其允许从简单SNP（其中在固定位置发生并且具有有限等位基数的变体）检测遗传变异与更复杂的序列变异模式（例如与多基因家族或生物体的多种遗传菌株一样，其中各个成员之间的序列变异既不固定也不一致）。阵列由连接到固体表面的短合成寡核苷酸探针组成，其与单链靶标杂交。可以使用使用在5'端修饰的引物来防止引入降解未保护链的5'-外切核酸酶降解的方法来产生单链靶标。本发明还提供用于将寡核苷酸探针固定到阵列表面的印刷缓冲液/溶液。本发明还提供杂交和洗涤缓冲液和条件以使杂交特异性和信号强度最大化，并减少杂交时间。

2. 发明授权

US08399197B2 Methods for amplifying polymeric nucleic acids 有权
公开(公告)号：US08399197B2
公开(公告)日：2013-03-19
申请号：US12555967
申请日：2009-09-09
申请人： Mark Aaron Behlke , Joseph Alan Walder , Jeffrey A. Manthey
发明人： Mark Aaron Behlke , Joseph Alan Walder , Jeffrey A. Manthey
IPC分类号： C12Q1/68 , C07H21/04
CPC分类号： C12Q1/686 , C12Q1/6848 , C12Q1/6853 , C12Q2527/143 , C12Q2525/101 , C12Q2549/119 , C12Q2525/186
摘要： The invention provides compositions and methods for amplifying nucleic acid polymer sequences in a high complexity nucleic acid sample. The unique compositions of the invention include a primer set composed of a mixture of two types of primers for DNA synthesis. For extension in one direction, the primers all contain modifications that destroy their ability to serve as templates that can be copied by DNA polymerases. For extension in the opposite direction the set includes at least one primer that can serve as a template and be replicated by DNA polymerases throughout its length. The method can be carried out by mixing the nucleic acid polymer sequence of interest with the set of DNA synthesis primers in an amplification reaction mixture. The reaction mixture is then subjected to temperature cycling analogous to the temperature cycling in PCR reactions. At least one primer in the primer set hybridizes to the nucleic acid polymer. It is preferred that the non-replicable primer hybridizes to the nucleic acid polymer and is extended to produce an extension product that contains sequence from the nucleic acid polymer to which the replicable primer then hybridizes. Of course, if the nucleic acid polymer is double stranded, both the replicable and nonreplicable primers will hybridize and be extended by DNA polymerase.

3. 发明申请

US20120123751A1 Method for Estimating a Melting Temperature of a Nucleic Acid in Buffers Containing Magnesium Ions 审中-公开
标题翻译：用于估计含有镁离子的缓冲液中核酸的熔融温度的方法
公开(公告)号：US20120123751A1
公开(公告)日：2012-05-17
申请号：US13290899
申请日：2011-11-07
申请人： Richard Owczarzy , Bernardo Moreira , Yong You , Mark Aaron Behlke , Joseph Alan Walder
发明人： Richard Owczarzy , Bernardo Moreira , Yong You , Mark Aaron Behlke , Joseph Alan Walder
IPC分类号： G06F19/12 , G06F17/10
CPC分类号： G16B25/00 , G16B20/00
摘要： The invention relates to methods and systems for predicting or estimating the melting temperature of duplex nucleic acids, in the presence of divalent cations, particularly duplexes of oligonucleotides which may be used as, for example, but not limited to primers or probes in PCR and/or hybridization assays. The methods and algorithms use novel formulas, having terms and coefficients that are functions of the particular nucleotide sequence, to estimate the effect of divalent cation salt conditions on the melting temperature.
摘要翻译：本发明涉及用于预测或估计双链核酸的解链温度的方法和系统，所述方法和系统在二价阳离子，特别是寡核苷酸的双链体存在下可用作例如但不限于PCR和/ 或杂交测定。方法和算法使用具有作为特定核苷酸序列的功能的术语和系数的新型公式来估计二价阳离子盐条件对熔融温度的影响。

4. 发明申请

US20100167353A1 RNASE H-BASED ASSAYS UTILIZING MODIFIED RNA MONOMERS 有权
标题翻译：使用修饰的RNA单体的RNASE H基测定
公开(公告)号：US20100167353A1
公开(公告)日：2010-07-01
申请号：US12507142
申请日：2009-07-22
申请人： Joseph Alan Walder , Mark Aaron Behlke , Scott Rose , Joseph Dobosy
发明人： Joseph Alan Walder , Mark Aaron Behlke , Scott Rose , Joseph Dobosy
IPC分类号： C12P19/34
CPC分类号： C12Q1/6848 , C12Q1/6844 , C12Q1/6853 , C12Q1/6858 , C12Q1/686 , C12Q2549/101 , C12Q2525/186 , C12Q2525/121 , C12Q2521/301
摘要： The present invention pertains to novel oligonucleotide compounds for use in various biological assays, such as nucleic acid amplification, ligation and sequencing reactions. The novel oligonucleotides comprise a ribonucleic acid domain and a blocking group at or near the 3′ end of the oligonucleotide. These compounds offer an added level of specificity previously unseen. Methods for performing nucleic acid amplification, ligation and sequencing are also provided. Additionally, kits containing the oligonucleotides are also disclosed herein.
摘要翻译：本发明涉及用于各种生物测定的新型寡核苷酸化合物，例如核酸扩增，连接和测序反应。该寡核苷酸在寡核苷酸的3'末端或其附近包含核糖核酸结构域和封闭基团。这些化合物提供了先前未见的特异性增加水平。还提供了进行核酸扩增，连接和测序的方法。此外，本文还公开了含有寡核苷酸的试剂盒。

5. 发明授权

US07439341B2 Fluorescence quenching azo dyes, their methods of preparation and use 有权
标题翻译：荧光猝灭偶氮染料，其制备和使用方法
公开(公告)号：US07439341B2
公开(公告)日：2008-10-21
申请号：US10987608
申请日：2004-11-12
申请人： Andrei Laikhter , Mark Aaron Behlke , Joseph Walder , Kevin William Roberts , Yawfui Yong
发明人： Andrei Laikhter , Mark Aaron Behlke , Joseph Walder , Kevin William Roberts , Yawfui Yong
IPC分类号： C09B29/01 , C09B29/08 , C07H21/04 , C12Q1/68
CPC分类号： C09B29/00 , C07C245/10 , C07F9/091 , C07F9/2408 , C07H21/04 , Y10T436/143333
摘要： Disclosed is a group of azo quencher compositions useful as fluorescence quenchers having the general structure of formula 1, methods of making or using the compositions, and kits comprising the composition.
摘要翻译：公开了一组可用作具有式1的通式结构的荧光猝灭剂的偶氮猝灭剂组合物，制备或使用该组合物的方法，以及包含该组合物的试剂盒。

6. 发明授权

US09434988B2 RNase H-based assays utilizing modified RNA monomers 有权
标题翻译：使用修饰的RNA单体的基于RNA酶H的测定
公开(公告)号：US09434988B2
公开(公告)日：2016-09-06
申请号：US13429077
申请日：2012-03-23
申请人： Mark Aaron Behlke , Scott D. Rose , Joseph Dobosy , Joseph Alan Walder
发明人： Mark Aaron Behlke , Scott D. Rose , Joseph Dobosy , Joseph Alan Walder
IPC分类号： C12Q1/68 , C12P19/34
CPC分类号： C12Q1/6858 , C12Q2521/327 , C12Q2563/107
摘要： The present invention provides methods of cleaving a nucleic acid strand to initiate, assist, monitor or perform biological assays.
摘要翻译：本发明提供了切割核酸链以启动，辅助，监测或进行生物测定的方法。

7. 发明授权

US08377641B2 Fluorescence quenching azo dyes, their methods of preparation and use 有权
标题翻译：荧光猝灭偶氮染料，其制备和使用方法
公开(公告)号：US08377641B2
公开(公告)日：2013-02-19
申请号：US13298479
申请日：2011-11-17
申请人： Andrei Laikhter , Mark Aaron Behlke , Joseph Walder , Kevin William Roberts , Yawfui Yong
发明人： Andrei Laikhter , Mark Aaron Behlke , Joseph Walder , Kevin William Roberts , Yawfui Yong
IPC分类号： C07H21/00 , C12Q1/68 , G01N33/52
CPC分类号： C09B29/00 , C07C245/10 , C07F9/091 , C07F9/2408 , C07H21/04 , Y10T436/143333
摘要： Disclosed is a group of azo quencher compositions useful as fluorescence quenchers having the general structure of formula 1, methods of making or using the compositions, and kits comprising the composition.
摘要翻译：公开了一组可用作具有式1的通式结构的荧光猝灭剂的偶氮猝灭剂组合物，制备或使用组合物的方法，以及包含该组合物的试剂盒。

8. 发明申请

US20120258455A1 RNase H-Based Assays Utilizing Modified RNA Monomers 有权
标题翻译：使用修饰的RNA单体的RNA酶H基测定法
公开(公告)号：US20120258455A1
公开(公告)日：2012-10-11
申请号：US13429077
申请日：2012-03-23
申请人： Mark Aaron Behlke , Scott D. Rose , Joseph Dobosy , Joseph Alan Walder
发明人： Mark Aaron Behlke , Scott D. Rose , Joseph Dobosy , Joseph Alan Walder
IPC分类号： C12P19/34 , G01N21/64 , G01N27/62 , C12Q1/68
CPC分类号： C12Q1/6858 , C12Q2521/327 , C12Q2563/107
摘要： The present invention provides methods of cleaving a nucleic acid strand to initiate, assist, monitor or perform biological assays.
摘要翻译：本发明提供了切割核酸链以启动，辅助，监测或进行生物测定的方法。

9. 发明授权

US08084588B2 Fluorescence quenching azo dyes, their methods of preparation and use 有权
标题翻译：荧光猝灭偶氮染料，其制备和使用方法
公开(公告)号：US08084588B2
公开(公告)日：2011-12-27
申请号：US12252721
申请日：2008-10-16
申请人： Andrei Laikhter , Mark Aaron Behlke , Joseph Walder , Kevin William Roberts , Yawfui Yong
发明人： Andrei Laikhter , Mark Aaron Behlke , Joseph Walder , Kevin William Roberts , Yawfui Yong
IPC分类号： C09B29/085 , C12Q1/68
CPC分类号： C09B29/00 , C07C245/10 , C07F9/091 , C07F9/2408 , C07H21/04 , Y10T436/143333
摘要： Disclosed is a group of azo quencher compositions useful as fluorescence quenchers having the general structure of formula 1, methods of making or using the compositions, and kits comprising the composition.
摘要翻译：公开了一组可用作具有式1的通式结构的荧光猝灭剂的偶氮猝灭剂组合物，制备或使用该组合物的方法，以及包含该组合物的试剂盒。

10. 发明授权

US08067164B2 Microarray system with improved sequence specificity 有权
标题翻译：具有改善序列特异性的微阵列系统
公开(公告)号：US08067164B2
公开(公告)日：2011-11-29
申请号：US12190446
申请日：2008-08-12
申请人： Kerry B. Gunning , Mark Aaron Behlke
发明人： Kerry B. Gunning , Mark Aaron Behlke
IPC分类号： C12Q1/68 , C12P19/34 , C07H21/02 , C07H19/04 , C40B40/06
CPC分类号： C12Q1/6837 , B01J19/0046 , B01J2219/0059 , B01J2219/00608 , B01J2219/00626 , B01J2219/00722 , C12P19/34 , C12Q1/6806 , Y10T436/108331 , C12Q2525/125 , C12Q2533/101 , C12Q2521/319
摘要： The invention provides a novel array method for nucleic acid sequence detection with improved specificity which allows for detection of genetic variation, from simple SNPs (where the variation occurs at a fixed position and is of limited allelic number) to more complex sequence variation patterns (such as with multigene families or multiple genetic strains of an organism where the sequence variation between the individual members is neither fixed nor consistent). The array is comprised of short, synthetic oligonucleotide probes attached to a solid surface which are hybridized to single-stranded targets. Single stranded targets can be produced using a method that employs primers modified on the 5′ end to prohibit degradation by a 5′-exonuclease that is introduced to degrade the unprotected strand. The invention further provides for printing buffers/solutions for the immobilization of oligonucleotide probes to an array surface. The invention also provides hybridization and wash buffers and conditions to maximize hybridization specificity and signal intensity, and reduce hybridization times.
摘要翻译：本发明提供了一种用于核酸序列检测的新型阵列方法，其具有改进的特异性，其允许从简单SNP（其中在固定位置发生并且具有有限等位基数的变体）检测遗传变异与更复杂的序列变异模式（例如与多基因家族或生物体的多种遗传菌株一样，其中各个成员之间的序列变异既不固定也不一致）。阵列由连接到固体表面的短合成寡核苷酸探针组成，其与单链靶标杂交。可以使用使用在5'端修饰的引物来防止引入降解未保护链的5'-外切核酸酶降解的方法来产生单链靶标。本发明还提供用于将寡核苷酸探针固定到阵列表面的印刷缓冲液/溶液。本发明还提供杂交和洗涤缓冲液和条件以使杂交特异性和信号强度最大化，并减少杂交时间。

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