基因组探针非试剂盒标记系统中DNaseI用量优化

Optimizing DNase I Dosage in Non-kit Labeling System of Genomic Probe

摘要: 用10 U的DNA Polym eraseⅠ,一个跨度达百万倍的DNaseⅠ量,在15℃分别标记了1μg甘蓝型油菜中油821的基因组DNA,发现DNaseⅠ用量为0.1 U时,10 m in后就可获得较好的探针标记效果.DNase I用量为0.01 U时,标记2 h就显示较深的颜色.随着DNaseⅠ用量的降低,标记时间需要相应地延长.其用量超过1 U时,无法获得较好的探针标记效果.综合考虑DNaseⅠ用量及标记时间,DNase I用量为0.01 U、标记时间为3~7 h获得的标记效果最佳.

Abstract: 1 μg genomic DNAs of Braasica napus L.Zhongyou 821 were labeled with 10U DNA Polymerase Ⅰ and a millionfold span of DNase Ⅰ dosage,respectively.A heavy dot blot can be displayed after 10 minutes with 0.1 U DNase Ⅰand after 2 hours with(0.01 U) DNase Ⅰ,respectively.The lower the DNase Ⅰdosage is used,the longer the labeling time is.A light one can only be displayed when the DNase Ⅰ dosage is over 1 U.The optimizing labeled probe can be obtained under 0.01U DNase I and 3~7 hours labeling time if we take these...