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 (Tm) 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.

中文翻译：

---

用于研究核酸变性的 Forster 共振能量转移：化学和生物科学实验室实验

Förster 共振能量转移 (FRET) 熔解分析旨在评估 DNA 二级结构的展开、变性过程及其使用称为 DNA 结合剂的化合物的稳定性，其中一些对 G-四链体 DNA 与双链体 DNA 具有高度特异性。首先，学生们确定了在没有 DNA 结合剂的情况下用 5'-FAM（荧光素）和 3'-TAMRA（四甲基罗丹明）双重标记的 DNA 序列的熔解温度 (Tm)。其次，他们通过监测荧光来确定 DNA 结合剂存在时 DNA 的熔解温度。完成这个实验后，学生们了解到这种方法可以进行半定量分析，以针对 DNA 二级结构测试各种 DNA 结合剂，