核糖核酸酶A在端粒免疫荧光原位杂交实验中的应用  

Application of RNase A in Telomere IF-FISH Experiment

作　　者：于芳[1] 金蕊[1] 黄君健[1] YU Fang;JIN Rui;HUANG Jun-Jian(Beijing Institute of Biotechnology, Beijing 100850, China)

机构地区：[1]军事医学科学院生物工程研究所,北京100850

出　　处：《生物技术通讯》2017年第5期681-684,共4页Letters in Biotechnology

摘　　要：目的:在端粒免疫荧光原位杂交实验中,降低核仁着色对端粒图像清晰度的影响。方法:分别将0.1、0.2、0.4 mg/m L的核糖核酸酶(RNase)A与待检标本于37℃消化过夜,然后进行端粒原位杂交实验。结果:0.4 mg/m L RNase A可以消除Hep G2细胞端粒原位杂交时的非特异结合,使用He La、293T和U2OS细胞也有同样效果。结论:在进行端粒原位杂交实验时,用0.4 mg/m L RNase A消化核仁可减少探针的非特异结合,提高端粒图像的清晰度。Objective:To decrease nucleolus coloring in immunofluorescence-fluorescence in situ hybridization(IF-FISH)of telomere.Methods:The cells were digested overnight at37℃with RNase A at concentration of0.1,0.2and0.4mg/mL and subjected to telomere in situ hybridization.Results:The use of0.4mg/mL RNase Aeliminates nonspecific binding of HepG2telomere in situ hybridization and ensure the accuracy of the experimental results.The same results were also got in HeLa,293T and U2OS cells.Conclusion:In situ hybridization oftelomere,digesting nucleoli with RNase A can provide a clearer immunofluorescence imaging and specificity.

关 键 词：核糖核酸酶A 免疫荧光原位杂交(IF-FISH) 端粒 

分 类 号：Q78[生物学—分子生物学]