CICS-UBI - Centro de Investigação em Ciências da Saúde, Universidade da Beira Interior, Covilhã, Portugal.
Biochem Mol Biol Educ. 2020 Jul;48(4):329-336. doi: 10.1002/bmb.21353. Epub 2020 Apr 8.
The Förster resonance energy transfer (FRET) melting assay intends to evaluate the unfolding, denaturation process of DNA secondary structures, and its stabilization using compounds known as DNA binders, some of which are highly specific for G-quadruplex DNAs versus duplex DNAs. First, students determined the melting temperature (T ) of DNA sequences double labeled with 5'-FAM (fluorescein) and 3'-TAMRA (tetramethylrhodamine) in the absence of DNA binders. Second, they determined the melting temperature of the DNAs in the presence of DNA binders by monitoring fluorescence. After completing this experiment, students understood that this method allows a semiquantitative analysis to test a variety of DNA binders against DNA secondary structures, and it can be used to rapidly identify the most promising drug candidates in the drug development stages at the basic research level.
Förster 共振能量转移 (FRET) 融解分析旨在评估 DNA 二级结构的解折叠、变性过程，以及使用被称为 DNA 结合物的化合物对其进行稳定化，其中一些 DNA 结合物对 G-四链体 DNA 比对双链 DNA 具有高度特异性。首先，学生们在没有 DNA 结合物的情况下测定了用 5'-FAM（荧光素）和 3'-TAMRA（四甲基罗丹明）双标记的 DNA 序列的融解温度 (T)。其次，他们通过监测荧光来测定 DNA 结合物存在时的 DNA 融解温度。完成这个实验后，学生们了解到，这种方法允许进行半定量分析，以测试各种 DNA 结合物对 DNA 二级结构的作用，并且可以用于在基础研究层面的药物开发阶段快速识别最有前途的候选药物。
