发明名称 |
Detection of nucleic acids by fluorescence quenching |
摘要 |
A detector oligonucleotide having a sequence which forms an intramolecularly base-paired secondary structure is described for use in detecting nucleic acid target sequences and target sequence amplification. The detector oligonucleotide is further modified by linkage to two dyes which form a donor/acceptor dye pair. The two dyes are positioned on the detector oligonucleotide such that they are in close spatial proximity in the base-paired, folded secondary structure, thereby causing quenching of donor fluorescence. The detector oligonucleotide may optionally further comprise a restriction endonuclease recognition site (RERS) which remains partially or entirely single-stranded in the base-paired secondary structure. The RERS is flanked by the two dyes. In the presence of target, the base-paired secondary structure is unfolded or linearized, increasing the distance between the donor and acceptor dyes and causing a change in fluorescence of the donor and/or the acceptor. If an RERS is present, it is rendered double-stranded in the presence of target, allowing cleavage or nicking by a restriction endonuclease and separation of the two dyes onto separate nucleic acid fragments. This may further contribute to the magnitude of the change in fluorescence.
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申请公布号 |
US5958700(A) |
申请公布日期 |
1999.09.28 |
申请号 |
US19990237510 |
申请日期 |
1999.01.26 |
申请人 |
BECTON, DICKINSON AND COMPANY |
发明人 |
NADEAU, JAMES G.;PITNER, J. BRUCE;LINN, C. PRESTON;SCHRAM, JAMES L. |
分类号 |
G01N33/50;C12N15/09;C12Q1/68;G01N33/566;(IPC1-7):C12Q1/68;C12P19/34;C07H21/04 |
主分类号 |
G01N33/50 |
代理机构 |
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代理人 |
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主权项 |
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地址 |
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