检测DNA杂交使用镧系荧光插入物,特异性结合双链DNA。

T. Nojima, Yasumitsu Kondoh, S. Takenaka, Teruhisa Ichihara, Makoto Takagi, Hideo Tashiro, Kazuko Matsumoto
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引用次数: 5

摘要

为了开发一种既不需要标记也不需要扩增活细胞核酸的DNA微阵列系统,我们开发了一种新的方法来检测和定量目标DNA与固定在固体表面的探针DNA杂交。该方法利用荧光插层剂:萘二亚胺衍生物携带两个荧光四齿β -二酮- eu3 +螯合物。该化合物选择性地与固定在塑料化验板上的双链DNA (dsDNA)结合。与单链DNA (ssDNA)结合的化合物的数量可以忽略不计。Eu3+的荧光强度与固定DNA的数量成正比,表明该化合物与dsDNA定量结合。因此,该方法不仅可以用于检测dsDNA,还可以用于测量固体表面上DNA的数量。
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Detection of DNA hybridization by use of a lanthanide fluorescent intercalator that specifically binds to double stranded DNA.
Toward development of a DNA microarray system in which neither labeling nor amplification of the nucleic acids from living cell is required, we have developed a new method for the detection and quantification of target DNA hybridized with probe DNA fixed on a solid surface. This method utilizes a fluorescent intercalator: naphthalene diimide derivative carrying two fluorescent tetradentate beta-diketone-Eu3+ chelates. This compound selectively binds to double stranded DNA (dsDNA) fixed on a plastic assay plate. The amount of the compound bound to single stranded DNA (ssDNA) is negligible. The fluorescent intensity of Eu3+ was in proportion to the amount of the fixed DNA, showing that the compound quantitatively binds to dsDNA. Therefore, this method can be used not only to detect dsDNA, but also to measure the amount of DNA on a solid surface.
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