A transfection reporter for the prevention of false-negative results in molecular beacon experiments.
A transfection reporter for the prevention of false-negative results in molecular beacon experiments.
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用于防止分子信标实验中出现假阴性结果的转染报告基因。
DOI：
10.1016/j.ab.2013.04.027
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发表时间：
2013
影响因子：
2.9
通讯作者：
Shigenori Iwai
中科院分区：
文献类型：
--
作者：
Tatsuya Toga;Isao Kuraoka;Akira Yasui;Shigenori Iwai
我们以前开发了一种分子信标型探针来检测细胞碱基切除修复中的链断裂，并发现荧光团/猝灭连接子中的磷酸二酯键被切割。该反应被应用于一种转染剂，该转染剂在连接到另一种类型的荧光团的连接物中含有未经修饰的磷酸二酯。用探针和报告基因共转染细胞后，分别用信号检测切口处和确认正确的转染。这种方法将有助于防止使用分子信标型探针进行实验时出现假阴性结果。
We previously developed a molecular beacon-type probe to detect the strand scission in cellular base excision repair and found that the phosphodiester linkages in the fluorophore/quencher linkers were cleaved. This reaction was applied to a transfection reporter, which contained the unmodified phosphodiester in the linker to another type of fluorophore. After cotransfection of cells with the probe and the reporter, the signals were used to detect the incision and to confirm the proper transfection, respectively. This method will contribute to the prevention of false-negative results in experiments using molecular beacon-type probes.