Tombler E R, Deutsch D G
State University of New York at Stony Brook.
Biotechniques. 1993 Dec;15(6):1060-4.
A spectrofluorometric assay to detect hybridization of oligodeoxynucleotides is described. Ethidium dimer, which yields significant fluorescence enhancement when intercalated between adjacent base pairs, was employed as the probe. Maximum enhancement of fluorescence occurred at equimolar ratios of p(dA)20.p(dT)20, while negligible fluorescence occurred for either of these chains when not in the presence of the other. Base-paired oligonucleotides at concentrations as low as 20 nM nucleotide could be detected, and the assay was linear over the tested range of 20 to 350 nM nucleotide. The method was used to measure the temperature-dependent melting and hybridization of two self-complementary heteropolymers and the hybridization of a self-annealing (dG)20 preparation.
本文描述了一种用于检测寡脱氧核苷酸杂交的荧光分光光度法。当嵌入相邻碱基对之间时会产生显著荧光增强的溴化乙锭二聚体被用作探针。在p(dA)20.p(dT)20等摩尔比时荧光增强最大，而当这两条链单独存在时荧光可忽略不计。浓度低至20 nM核苷酸的碱基配对寡核苷酸都能被检测到，并且该检测方法在20至350 nM核苷酸的测试范围内呈线性。该方法用于测量两种自互补杂聚物的温度依赖性解链和杂交以及一种自退火(dG)20制剂的杂交。
