Kosman Joanna, Wu Yu-Tang, Gluszynska Agata, Juskowiak Bernard
Faculty of Chemistry, Adam Mickiewicz University in Poznan, Umultowska 89b, 61-614, Poznan, Poland,
Anal Bioanal Chem. 2014 Nov;406(28):7049-57. doi: 10.1007/s00216-014-8119-7. Epub 2014 Sep 12.
Characterization and optimization studies of N-methyl-4-hydrazino-7-nitrobenzofurazan (MNBDH) as a new fluorogenic substrate in the peroxidation reaction catalyzed by DNAzyme are reported. The effects of pH, H2O2 concentration, metal-cation type, and the concentration and type of surfactant on the fluorescence intensity were investigated. The optimized reaction was subsequently used for the development of an assay for DNA detection based on a molecular-beacon probe. The use of a fluorogenic substrate enabled the detection of a single-stranded DNA target with a 1 nmol L(-1) detection limit.
报道了N-甲基-4-肼基-7-硝基苯并呋喃(MNBDH)作为DNAzyme催化过氧化反应中一种新型荧光底物的表征和优化研究。研究了pH、H2O2浓度、金属阳离子类型以及表面活性剂的浓度和类型对荧光强度的影响。随后,将优化后的反应用于基于分子信标探针的DNA检测分析方法的开发。使用荧光底物能够检测单链DNA靶标,检测限为1 nmol L(-1)。