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11.发明授权公开(公告)号：US08530194B2
公开(公告)日：2013-09-10
申请号：US12088213
申请日：2006-09-26
CPC分类号： C12Q1/6848 , C12Q2527/101 , C12Q2525/205
摘要： Aspects of the invention relate to the use of novel oligonucleotides as temperature-sensitive inhibitors for thermostable DNA polymerases. Some inhibitors exhibit temperature-dependent and, in some cases, reversible inhibitory property by changing the conformation of at least a portion of the oligonucleotides from double-stranded to single stranded or in some cases vice versa in a temperature-dependent manner. Aspects also relate to the use of an the inhibitors in a hot-start PCR compositions, wherein the inhibitor may act to suppress the activity of the thermostable DNA polymerase below a desired activation temperature, Tact, and wherein the inhibitor is thermally inactivated above Tact, thus liberating the polymerase activity and initiating the DNA amplification process. Aspects further relate to a procedure for formulating the composition of a hot-start PCR reaction mixture. The hot-start PCR methods disclosed herein are generally faster, more flexible and lower in cost than existing methods.
摘要翻译： 本发明的方面涉及新型寡核苷酸作为热稳定性DNA聚合酶的温度敏感性抑制剂的用途。 一些抑制剂在温度依赖性方面通过将寡核苷酸的至少一部分构型从双链改变为单链或在一些情况下反之亦然地表现出温度依赖性，并且在一些情况下是可逆抑制性质。 方面还涉及在热启动PCR组合物中使用抑制剂，其中抑制剂可以起作用以将热稳定性DNA聚合酶的活性抑制在所需活化温度Tact以下，并且其中抑制剂在Tact之上热灭活， 从而释放聚合酶活性并引发DNA扩增过程。 方面还涉及用于配制热启动PCR反应混合物的组合物的方法。 本文公开的热启动PCR方法通常比现有方法更快，更灵活并且成本更低。
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公开(公告)号：US07667024B2
公开(公告)日：2010-02-23
申请号：US10993625
申请日：2004-11-19
CPC分类号： C12Q1/6818 , C12Q1/6851 , Y10S435/975 , C12Q2565/1015 , C12Q2561/101 , C12Q2565/1025
摘要： An embodiment of the invention discloses new methods for designing labeled nucleic acid probes and primers by labeling oligonucleotides with a plurality of spectrally identical or similar dyes and optionally with one or more quencher dyes. Oligonucleotides labeled in accordance with some embodiments of the invention exhibit a detectable increase in signal, for example, fluorescent signal when the labeling dyes are separated from one another. Methods for separating the dye include cleaving the labeled oligonucleotides include using enzymes that have 5′-exonuclease activity. In one embodiment nucleic acid primers of the present invention may fluoresce upon hybridization to a target sequence and incorporation into the amplification product. Nucleic acid probes and primers of the present invention have wide applications ranging from general detection of a target nucleic acid sequence to clinical diagnostics. Major advantages of the oligonucleotides including nucleic acid probes and primers of many embodiments of the present invention are their synthetic simplicity, spectral versatility and superior fluorescent signal.
摘要翻译： 本发明的一个实施方案公开了通过用多个光谱相同或相似的染料和任选地与一种或多种猝灭剂染料标记寡核苷酸来设计标记的核酸探针和引物的新方法。 根据本发明的一些实施方案标记的寡核苷酸在标记染料彼此分离时表现出可检测的信号增加，例如荧光信号。 用于分离染料的方法包括切割标记的寡核苷酸包括使用具有5'-外切核酸酶活性的酶。 在一个实施方案中，本发明的核酸引物可以在与靶序列杂交时发荧光，并掺入扩增产物。 本发明的核酸探针和引物具有广泛的应用范围，从靶核酸序列的一般检测到临床诊断。 包括本发明许多实施方案的核酸探针和引物的寡核苷酸的主要优点是它们的合成简便性，光谱通用性和优异的荧光信号。
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13.发明申请公开(公告)号：US20090047714A1
公开(公告)日：2009-02-19
申请号：US12088213
申请日：2006-09-26
CPC分类号： C12Q1/6848 , C12Q2527/101 , C12Q2525/205
摘要： Aspects of the invention relate to the use of novel oligonucleotides as temperature-sensitive inhibitors for thermostable DNA polymerases. Some inhibitors exhibit temperature-dependent and, in some cases, reversible inhibitory property by changing the conformation of at least a portion of the oligonucleotides from double-stranded to single stranded or in some cases vice versa in a temperature-dependent manner. Aspects also relate to the use of an the inhibitors in a hot-start PCR compositions, wherein the inhibitor may act to suppress the activity of the thermostable DNA polymerase below a desired activation temperature, Tact, and wherein the inhibitor is thermally inactivated above Tact, thus liberating the polymerase activity and initiating the DNA amplification process. Aspects further relate to a procedure for formulating the composition of a hot-start PCR reaction mixture. The hot-start PCR methods disclosed herein are generally faster, more flexible and lower in cost than existing methods.
摘要翻译： 本发明的方面涉及新型寡核苷酸作为热稳定性DNA聚合酶的温度敏感性抑制剂的用途。 一些抑制剂在温度依赖性方面通过将寡核苷酸的至少一部分构型从双链改变为单链或在一些情况下反之亦然地表现出温度依赖性，并且在一些情况下是可逆抑制性质。 方面还涉及在热启动PCR组合物中使用抑制剂，其中抑制剂可以起作用以将热稳定性DNA聚合酶的活性抑制在所需活化温度Tact以下，并且其中抑制剂在Tact之上热灭活， 从而释放聚合酶活性并引发DNA扩增过程。 方面还涉及用于配制热启动PCR反应混合物的组合物的方法。 本文公开的热启动PCR方法通常比现有方法更快，更灵活并且成本更低。
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公开(公告)号：US20100285468A1
公开(公告)日：2010-11-11
申请号：US12679534
申请日：2008-09-24
申请人： Xing Xin
发明人： Xing Xin
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6883 , C12Q1/6827 , C12Q1/6886 , C12Q2600/156 , C12Q2565/632 , C12Q2563/173 , C12Q2545/101 , C12Q2531/113 , C12Q2527/107
摘要： The present invention provides compositions, methods, and kits for nucleic acids analyses. In particular, melting analyses are used to detect the presence or absence and to quantify nucleic acids.
摘要翻译： 本发明提供了用于核酸分析的组合物，方法和试剂盒。 特别地，使用熔解分析来检测核酸的存在或不存在以及定量。
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