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    • 89. 发明公开
    • PO절단 및 혼성화에 의한 타겟 핵산서열의 검출
    • 通过PO CLEAVAGE和杂交检测目标核酸序列
    • KR1020130137703A
    • 2013-12-17
    • KR1020137029140
    • 2012-05-03
    • 주식회사 씨젠
    • 천종윤이영조
    • C12Q1/68C12N15/10C12N15/11
    • C12Q1/6823C12Q1/6837C12Q1/6813C12N15/11C12Q2565/50C12Q2521/319C12Q2565/519
    • The present invention relates to detection of target nucleic acid sequences on a solid phase substrate by POCH (PO cleavage and hybridization) assaies. The present invention detects target nucleic acid sequences using a fact that PO (probing oligonucleotide), which is hybridized with the target nucleic acid sequences, is amputated and the amputation of the PO is detected by the hybridization with CO (capturing oligonucleotide). In the present invention, the non-amputated PO is hybridized with a CO fixed on the solid phase substrate. In the present invention, designing of the PO and CO is convenient and optimization of a reaction condition is routinely facilitated. If the amputation of POs and detection of signals on the solid phase substrate are repeated and successively performed, the number of amputated POs is increased by the number of repetition of the amputative reactions and the signals are changed by the number of the amputated POs. Therefore, the target nucleic acid sequence can be detected in real time. While, if the target nucleic acid sequence is not existed, change of signals is not observed. [Reference numerals] (A) Hybridization;(AA) Upstream primer;(B) Primer extension & PO cutting;(BB) DNA or nucleic acid mixture;(C) Hybridization of CO and PO & detection;(CC) Downstream primer;(DD,FF) Cut PO;(EE,GG) Uncut PO;(HH) Reporter-contained piece;(II) Solid phase supporter;(R) Reporter
    • 本发明涉及通过POCH(PO切割和杂交)试验检测固相底物上的靶核酸序列。 本发明使用与靶核酸序列杂交的PO(探测寡核苷酸)截留并通过与CO(捕获寡核苷酸)的杂交来检测PO的截断的事实来检测靶核酸序列。 在本发明中,非截留的PO与固相在固相基板上的CO杂交。 在本发明中,PO和CO的设计是方便的,并且常规地促进了反应条件的优化。 如果重复并依次进行POs的截肢和对固相基板上的信号的检测,则截断的PO的数量增加了截肢反应的重复次数,并且信号被截断的PO的数目改变。 因此,可以实时检测靶核酸序列。 而如果目标核酸序列不存在,则不能观察到信号的改变。 (A)杂交;(AA)上游引物;(B)引物延伸和PO切割;(BB)DNA或核酸混合物;(C)CO和PO和检测的杂交;(CC)下游引物; (DD,FF)切割PO;(EE,GG)未切割PO;(HH)报告人包含的部分;(II)固相支持者;(R)记者