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Nucleic acid amplification oligonucleotides with molecular energy transfer labels and methods based thereon

机译:具有分子能量转移标记的核酸扩增寡核苷酸及其基于方法

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.
机译:本发明提供了标记的核酸扩增寡核苷酸,其可以是线性或发夹引物或封闭寡核苷酸。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量被受体吸收。受体可以是在不同于供体部分的波长下发荧光的荧光团,或者可以是猝灭剂。配置本发明的寡核苷酸,使得供体部分和受体部分被掺入扩增产物中。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,用核酸外切酶或使用特定温度进行处理,无需分离未结合的引物。这种“封闭管”形式大大降低了扩增产物对残留物污染的可能性,提供了高通量的样品,并且可以完全自动化。

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