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1.发明授权Nucleic acid amplification oligonucleotides with molecular energy transfer labels and methods based thereon 失效具有分子能量转移标记的核酸扩增寡核苷酸和基于其的方法
公开(公告)号：US6090552A
公开(公告)日：2000-07-18
申请号：US891516
申请日：1997-07-11
CPC分类号： C12Q1/6818 , C12Q1/6844 , C12Q1/6858 , C12Q1/686
摘要： The present invention provides labeled nucleic acid amplification oligonucleotides, which can be linear or hairpin primers or blocking oligonucleotides. The oligonucleotides of the invention are labeled with donor and/or acceptor moieties of molecular energy transfer pairs. The moieties can be fluorophores, such that fluorescent energy emitted by the donor is absorbed by the acceptor. The acceptor may be a fluorophore that fluoresces at a wavelength different from the donor moiety, or it may be a quencher. The oligonucleotides of the invention are configured so that a donor moiety and an acceptor moiety are incorporated into the amplification product. The invention also provides methods and kits for directly detecting amplification products employing the nucleic acid amplification primers. When labeled linear primers are used, treatment with exonuclease or by using specific temperature eliminates the need for separation of unincorporated primers. This "closed-tube" format greatly reduces the possibility of carryover contamination with amplification products, provides for high throughput of samples, and may be totally automated.
摘要翻译： 本发明提供标记的核酸扩增寡核苷酸，其可以是线性或发夹引物或阻断寡核苷酸。 本发明的寡核苷酸用分子能量转移对的供体和/或受体部分标记。 这些部分可以是荧光团，使得供体发射的荧光能被受体吸收。 受体可以是在与供体部分不同的波长处发荧光的荧光团，或者它可以是猝灭剂。 配置本发明的寡核苷酸使得供体部分和受体部分被并入扩增产物。 本发明还提供了使用核酸扩增引物直接检测扩增产物的方法和试剂盒。 当使用标记的线性引物时，用外切核酸酶处理或通过使用特定温度消除了对未引入引物的分离的需要。 这种“封闭管”格式大大降低了放大产品携带污染的可能性，提供了高通量的样品，并且可以完全自动化。
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公开(公告)号：US5866336A
公开(公告)日：1999-02-02
申请号：US778487
申请日：1997-01-03
CPC分类号： C12Q1/686 , C12Q1/6818 , C12Q1/6844
摘要： The present invention provides labeled nucleic acid amplification oligonucleotides, which can be linear or hairpin primers or blocking oligonucleotides. The oligonucleotides of the invention are labeled with donor and/or acceptor moieties of molecular energy transfer pairs. The moieties can be fluorophores, such that fluorescent energy emitted by the donor is absorbed by the acceptor. The acceptor may be a fluorophore that fluoresces at a wavelength different from the donor moiety, or it may be a quencher. The oligonucleotides of the invention are configured so that a donor moiety and an acceptor moiety are incorporated into the amplification product. The invention also provides methods and kits for directly detecting amplification products employing the nucleic acid amplification primers. When labeled linear primers are used, treatment with exonuclease or by using specific temperature eliminates the need for separation of unincorporated primers. This "closed-tube" format greatly reduces the possibility of carryover contamination with amplification products, provides for high throughput of samples, and may be totally automated.
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公开(公告)号：US5728526A
公开(公告)日：1998-03-17
申请号：US472239
申请日：1995-06-07
CPC分类号： C12Q1/6834 , C12Q2600/156
摘要： A method for analyzing a target nucleotide sequence which exists in a first state or a different second state which makes the method particularly useful for determining point mutations. The method uses a first polynucleotide which is immobilized on a solid support and which is at least partially complementary to a first segment of the target nucleotide sequence. By means of a series of steps, a product of the first polynucleotide and a further polynucleotide that contains a detectable label can be obtained. When the state to be analyzed occurs in a rare population, amplification can be conducted so that substantially only amplification of the target nucleotide sequence in one of the states is attained. The method can be used to analyze multiple target sequences simultaneously. A kit which can be used in the method is also set forth.
摘要翻译： 用于分析以第一状态或不同第二状态存在的靶核苷酸序列的方法，其使得该方法对于确定点突变特别有用。 该方法使用固定在固体支持物上并且与靶核苷酸序列的第一片段至少部分互补的第一多核苷酸。 通过一系列步骤，可以获得含有可检测标记的第一多核苷酸和另外的多核苷酸的产物。 当要分析的状态发生在罕见的群体中时，可以进行扩增，使得仅在一种状态下实现目标核苷酸序列的扩增。 该方法可用于同时分析多个靶序列。 还列出了可用于该方法的试剂盒。
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公开(公告)号：US6117635A
公开(公告)日：2000-09-12
申请号：US837034
申请日：1997-04-11
CPC分类号： C12Q1/6818 , C12Q1/6844 , C12Q1/6858 , C12Q1/686
摘要： The present invention provides labeled nucleic acid amplification oligonucleotides, which can be linear or hairpin primers or blocking oligonucleotides. The oligonucleotides of the invention are labeled with donor and/or acceptor moieties of molecular energy transfer pairs. The moieties can be fluorophores, such that fluorescent energy emitted by the donor is absorbed by the acceptor. The acceptor may be a fluorophore that fluoresces at a wavelength different from the donor moiety, or it may be a quencher. The oligonucleotides of the invention are configured so that a donor moiety and an acceptor moiety are incorporated into the amplification product. The invention also provides methods and kits for directly detecting amplification products employing the nucleic acid amplification primers. When labeled linear primers are used, treatment with exonuclease or by using specific temperature eliminates the need for separation of unincorporated primers. This "closed-tube" format greatly reduces the possibility of carryover contamination with amplification products, provides for high throughput of samples, and may be totally automated.
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公开(公告)号：US5593840A
公开(公告)日：1997-01-14
申请号：US461823
申请日：1995-06-05
CPC分类号： C12Q1/6848 , C12Q1/6844 , C12Q1/6853 , C12Q1/686 , C12Q1/6862 , C12Q1/6876 , C12Q2600/156
摘要： A process for amplifying nucleic acid sequences from a DNA or RNA template which may be purified, or may exist in a mixture of nucleic acids. The resulting nucleic acid sequences may be exact copies of the template, or may be modified. The process has advantages over prior art amplification processes in that it increases the fidelity of copying a specific nucleic acid sequence, and it allows one to more efficiently detect a particular point mutation in a single assay.
摘要翻译： 从DNA或RNA模板扩增核酸序列的方法，该DNA或RNA模板可被纯化，或可存在于核酸混合物中。 所得到的核酸序列可以是模板的精确拷贝，或可以被修饰。 该方法具有优于现有技术扩增方法的优点，因为它增加复制特定核酸序列的保真度，并且其允许在单个测定中更有效地检测特定的点突变。
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公开(公告)号：US06835537B1
公开(公告)日：2004-12-28
申请号：US09721918
申请日：2000-11-27
IPC分类号： C12Q168
CPC分类号： C12Q1/6869 , C12Q2525/101
摘要： A method for normalizing the intensity of G bands in sequencing methods which utilize dITP is presented. The use of dITP normally results in decreased intensities of G bands which occur after A bands, i.e., in the sequence AG. It has been discovered that the use of ddITP in place of ddGTP or in conjunction with ddGTP helps to normalize the intensity of the G bands following A bands. This aids in preventing errors in reading sequencing chromatograms and allows for extended reads of sequencing chromatograms as compared to methods which utilize dITP without the use of ddITP.
摘要翻译： 提出了一种利用dITP的测序方法对G带强度进行归一化的方法。 dITP的使用通常导致在A条带之后，即在序列AG中发生的G带的强度降低。 已经发现，使用ddITP代替ddGTP或与ddGTP结合有助于使A带之后的G带的强度标准化。 与使用dITP而不使用ddITP的方法相比，这有助于预测读取测序色谱图中的错误，并允许扩展读取测序色谱图。
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