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核酸分子杂交法广泛用于分子生物学研究,对遗传性疾病的探索、优生检查、法医侦破罪犯等都起到重要作用。在分子杂交中,DNA探针的制备是方法成功的关键。目前常用的是~(32)P或~(35)S标记的探针,但在国内高比活性的底物[α-~(32)P]-dATP或[α-~(35)S]-dATP都难得到,阻碍了分子杂交法的应用。 1981年Langer用dATP-生物素做底物,标记DNA,再与过氧化酶联的抗生物素结合,经酶反应显色检测。1983年Leary用碱性磷酸酶代替过氧化物酶,使检测的灵敏度提高了20～50倍,达到同
Hybridization of nucleic acid molecules is widely used in molecular biology research, genetic disease exploration, eugenics check, forensic detection of criminals have played an important role. In molecular hybridization, the preparation of DNA probes is the key to the success of the method. At present, the probe of ~ (32) P or ~ (35) S is commonly used, but the high specific activity of [α- (32) P] -dATP or [α- ~ (35) S] -dATP are hard to get, hinder the application of molecular hybridization method. In 1981, Langer used dATP-biotin as a substrate, labeled DNA, and then conjugated with peroxidase biotin, and detected by enzyme reaction. 1983 Leary alkaline phosphatase instead of peroxidase, the detection sensitivity increased by 20 to 50 times, to achieve the same