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2. 发明公开

EP3174998A1 METHOD FOR SELECTING A TARGET NUCLEIC ACID SEQUENCE 有权转让
标题翻译：用于选择目标核酸序列的方法
公开(公告)号：EP3174998A1
公开(公告)日：2017-06-07
申请号：EP15744928.1
申请日：2015-07-31
申请人： Olink Bioscience AB
发明人： LANDEGREN, Ulf , NONG, Yuan
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6806 , C12Q1/6853 , C12Q2521/501 , C12Q2525/131 , C12Q2525/161 , C12Q2525/301 , C12Q2531/125 , C12Q2533/101 , C12Q2533/107
摘要： 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.
摘要翻译：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。

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