“高效RAPD”技术的初步研究及其PAGE检测  

Preliminary studies on "High Efficient RAPD" technique and its PAGE detection

作　　者：孟宪红[1] 孔杰[1] 刘萍[1] 

机构地区：[1]中国水产科学研究院黄海水产研究所,青岛266071

出　　处：《海洋水产研究》2004年第2期13-17,共5页Marine Fisheries Research

基　　金：国家高技术研究发展计划 (863计划 ) (2 0 0 3AA60 30 2 1 );国家重点基础研究发展规划项目 (G1 9990 1 2 0 0 7);农业部海洋渔业资源可持续利用重点开放实验室开放课题 (2 0 0 2 0 7)共同资助

摘　　要：对经典的单引物RAPD技术进行优化 ,确立了适合于多条引物 (2～ 4条 )同时参与同一反应的“高效RAPD”体系及扩增运行条件 ,不但实现了 1次操作检测更多位点、使多样本量RAPD检测变得相对省时省力 ,同时还可节省珍贵的实验样品。本文采用了具有高分辨能力的非变性PAGE 银染检测方法对RAPD产物进行检测 ,使一般经琼脂糖电泳检测仅能分辨出不足 10条谱带的RAPD产物可分辨出 15～ 30条清晰可见谱带。非变性PAGE 银染检测方法操作安全 ,银染后的凝胶可长期保存。The paper focused on the 'high efficient RAPD' technique, which means there is more than one primer (commonly 2～4 primers) in a reaction system. The reaction condition of 'high efficient RAPD' was established after optimizing the PCR factors based on the traditional simple primer RAPD. Compared to the traditional RAPD technique, 'high efficient RAPD' is more simple, less time and money consuming, as well as using very few of rare DNA as template. We can detect more loci in a single analysis by 'high efficient RAPD', at the same time, silver-stained & nondenatural PAGE electrophoresis was used to separated the 'high efficient RAPD' products instead of agarose & EB-stained electrophoresis, 15～30 bands of amplified products can be detected on the polyacrylamide gel while less than 10 bands on agarose gel. Furthermore, the method of silver-stained & nondenatural PAGE electrophoresis was safe and silver-stained gel can be saved for long time.

关 键 词：高效RAPD PAGE 银染 

分 类 号：Q953.1[生物学—动物学] Q78