定量荧光原位杂交测定端粒长度  

Telomere Length Measurement by Quantitative FISH

作　　者：曾思聪[1] 卢光琇[1] 

机构地区：[1]中南大学生殖与干细胞工程研究所人类干细胞国家工程研究中心,湖南长沙410078

出　　处：《激光生物学报》2011年第3期394-397,共4页Acta Laser Biology Sinica

基　　金：国家重点基础研究发展计划(973计划)(2007CB948103);中国高技术发展研究计划(863计划)(2006AA02A102);湖南省科技计划项目(2008FJ3 133)

摘　　要：目的:应用定量荧光原位杂交(Q-FISH)方法测定端粒长度。方法:选取4种端粒长度均一的标准细胞株采用Q-FISH的方法做出荧光亮度与端粒长度的标准曲线,从而得出实验细胞株的端粒长度,与DNA印迹法测定末端限制性片段(TRF)长度进行二者之间的相关性分析。结果:检测荧光强度的最佳线性曝光时间为400ms,相对于DNA印迹法,定量荧光原位杂交(Q-FISH)法所需标本量少,实验周期短,端粒长度结果与Southern杂交法具有很好的相关性。结论:采用定量荧光原位杂交方法测端粒长度具有重复性好、精确可靠的特点,适用于对珍贵标本的端粒改变进行分析。Objective： Our purpose of this study was to set up quantitative florescence suit hybridization （Q-FISH） to measure telomere length. Methods：4 cell lines which have homogeneous telomere length were selected to establish telomere length standard curve. Telomere length of GM847 ceils was evaluated according to the standard curve by Q-FISH or by telomere restriction fragment assay （TRF）. Relevance analysis was performed between Q-FISH and TRF experiment group. Results：Our experiments suggest that the best exposure time for collection signal in Q-FISH is 400 ms. Compared with TRF assay, Q-FISH needs less cell number and is time saving. Conclusions： Our results shows evaluating telomere length by Q-FISH has good reproducibility and is accurate than TRF assay, this method is particularly suitable for small size and rare sample analysis.

关 键 词：定量荧光原位杂交 端粒 

分 类 号：Q291[生物学—细胞生物学] Q343.242