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AFLP荧光标记和银染技术的比较分析
引用本文:赵翀,刘法央,郭玉霞,严学兵. AFLP荧光标记和银染技术的比较分析[J]. 甘肃农业大学学报, 2007, 42(2): 125-129
作者姓名:赵翀  刘法央  郭玉霞  严学兵
作者单位: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,中国上海,200433;郑州牧业工程高等专科学校,河南,郑州,450008
基金项目:科技部国际科技合作项目 , 河南省科技厅科技攻关项目 , 科技部合作项目
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.

关 键 词:AFLP  披碱草  扩增图谱  DNA多态性  银染法  荧光标记
文章编号:1003-4315(2007)02-0125-05
修稿时间:2006-10-11

Comparison between fluorescence labeling and silver-staining techniques of AFLP
ZHAO Chong,LIU Fa-yang,GUO Yu-xia,YAN Xue-bing. Comparison between fluorescence labeling and silver-staining techniques of AFLP[J]. Journal of Gansu Agricultural University, 2007, 42(2): 125-129
Authors:ZHAO Chong  LIU Fa-yang  GUO Yu-xia  YAN Xue-bing
Affiliation:1. College of LifeSciences and Teehnology,Gansu Agricultural University,Lanzhou 730070,China; 2. Engineering College of Animal Husbandry and Veterinary, Henan Agricultural University,Zhengzhou 450002,China; 3. Institute of Biodiversity Seienee,Fudan Unlverslty,Shanghai 200433,China; 4. Zhengzhou College of Animal Husbandry,Zhengzhou 450008,China
Abstract:The present study was intended to comparison of fluorescence labeling and silver-staining detection system of AFLP of Elymus species.Genomic DNA of Elymus 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.
Keywords:AFLP  Elymus  amplification map  DNA polymorphism  silver-staining  fluorescence labeling
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