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    • 2. 发明公开
    • METHOD FOR SELECTING A TARGET NUCLEIC ACID SEQUENCE
    • 用于选择目标核酸序列的方法
    • EP3174998A1
    • 2017-06-07
    • EP15744928.1
    • 2015-07-31
    • Olink Bioscience AB
    • LANDEGREN, UlfNONG, Yuan
    • C12Q1/68
    • C12Q1/6806C12Q1/6853C12Q2521/501C12Q2525/131C12Q2525/161C12Q2525/301C12Q2531/125C12Q2533/101C12Q2533/107
    • The present invention relates to a method for selecting a target region of interest (ROI) in a target nucleic acid molecule using a nucleic acid probe comprising a 3' sequence capable of hybridising to a target nucleic acid molecule and acting as a primer for the production of a complement of the target ROI (i.e. by target templated extension of the primer), and a sequence capable of templating the circularisation and ligation of the extended probe comprising the reverse complement of the target ROI and a portion of the probe. The circularised molecule thus obtained contains the reverse complement of the target ROI and may be subjected to further analysis and/or amplification etc. The probe may be provided as an oligonucleotide comprising a stem-loop structure or as a partially double- stranded construct and comprises a single-stranded 3' end region containing the target- binding site. A second binding site provided in the probe serves as the ligation template for circularisation, and the stem-loop structure, if present, is cleaved to render the second binding site available for hybridisation to the target complement. Also provided are probes and kits for carrying out such a method.
    • 本发明涉及使用包含能够与靶核酸分子杂交的3'序列并充当产生的引物的核酸探针选择靶核酸分子中的目标靶区域(ROI)的方法 (即通过引物的目标模板延伸),以及能够模板扩增探针的环化和连接的序列,所述扩增探针包含目标ROI和部分探针的反向互补物。 如此获得的环化分子含有目标ROI的反向互补物并且可以进行进一步分析和/或扩增等。探针可以作为包含茎环结构或部分双链构建体的寡核苷酸提供,并且包含 含有靶结合位点的单链3'端区域。 探针中提供的第二结合位点用作环化的连接模板,并且茎环结构(如果存在的话)被切割以使得第二结合位点可用于与靶补体杂交。 还提供了用于实施这种方法的探针和试剂盒。