AFLP荧光标记和银染技术的比较分析  

Comparison between fluorescence labeling and silver-staining techniques of AFLP

作　　者：赵翀[1] 刘法央[1] 郭玉霞[2] 严学兵[3] 

机构地区：[1]甘肃农业大学生命科学院,甘肃兰州730070 [2]河南农业大学牧医工程学院,河南郑州450002 [3]复旦大学生物多样性与生态工程教育部重点试验室

出　　处：《甘肃农业大学学报》2007年第2期125-129,共5页Journal of Gansu Agricultural University

基　　金：科技部国际合作项目(CB711103);河南省科技厅项目(0611032300);科技部合作项目(B711103).

摘　　要：为了建立准确、高效、实用的AFLP体系和方法,采用改进的CTAB-DNA提取方法从披碱草的嫩叶中提取降解少、样品纯度高、蛋白去除干净、不含PCR反应抑制剂及其他酶反应抑制剂的总DNA.选取3对引物扩增,采用AFLP荧光标记和银染法对披碱草全基因组多态性进行了分析,其中荧光标记法得到231条条带,银染法得到75条条带,平均每条引物荧光标记技术比银染法多得到52条条带,多态比率是银染法的1.8倍.据此认为荧光标记法检测效果较为理想,适于进行披碱草遗传多样性的分析和研究.The present study was intended to comparison of fluorescence labeling and silver-staining detection system of AFLP of Elyrnus species. Genomic DNA of Elyrnus nutans were abstracted from delicate leaves by means of CTAB. Three pairs of primers were used for amplification of genomic DNA, 231 bands were got by fluorescence labeling technique and 75 bands by silver-staining technique. The results showed that, compared with silver-staining technique, using fluorescence labeling could get much more bands in every site and better checked-effect. In addition, fluorescence labeling technique was more suitable for analyzing or studying descendible polymorphism than silver-staining technique. Analyzed the cost and work efficiency,with almost equal cost, the efficiency of fluorescent system was 1.8 times than that of silver-staining system without consideration of instrument investment. Some other strategies, such as collection of data,analysis of feasibility and data analysis using Sega procedure were also discussed.

关 键 词：AFLP 披碱草 扩增图谱 DNA多态性 银染法 荧光标记 

分 类 号：S819[农业科学—畜牧学]