生物素标记核酸探针杂交检测灵敏度的比较研究

STUDY ON THE DETECTING SENSITIVITY OF DNA HYBRIDIZED WITH BIOTINYLATED PROBES

用Biotin-11-dUTP进行缺口翻译制备生物素标记的人β-珠蛋白基因探针并用该探针进行了斑点杂交,硝酸纤维素膜印迹杂交及琼脂糖凝胶直接杂交方法学比较结果表明:经ABAP法显色后,上述三种杂交法中,DNA最低可检测量依次为5、10、50pg。

A biotin-labeled probe of β-globin gene was prepared by a nick-translation procedure in which the labeled deoxynucleotide resoure was biotin-11-dUTP. Using the biotin-labeled probe of β-globin gene, a comparative study was made for dot-blot, southern blot and direct gel hybridization method. The results showed that the minimum detectable amounts of target DNA in samples were 5, 10, and 50pg respectively. Dot-blot hybridization method was shown to be the most sensititive.