大菱鲆AFLP分析体系的建立  

The establishment of AFLP analysis system for turbot (Scophthalmus maximus)

作　　者：季士治[1] 王伟继[2] 雷霁霖[2] 孔杰[2] 

机构地区：[1]中国海洋大学生命科学与技术学部,青岛266003 [2]农业部海洋渔业资源可持续利用重点开放实验室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》2007年第1期6-12,共7页Marine Fisheries Research

基　　金：国家高技术研究发展863计划"海水养殖种子工程北方基地"课题资助

摘　　要：本研究构建了大菱鲆扩增片段长度多态性(AFLP)分析体系,对DNA提取、双酶切反应、连接反应、预扩增反应、选择性扩增反应和银染等步骤进行了分析。结果表明,用EcoRI/MseI双酶切、核心序列加3个选择性碱基的选择性扩增引物、1∶6(w∶w)的EcoRI端与MseI端选择性扩增引物比和20μl选择性扩增体积,可得到十分稳定的结果。所得产物经电泳和银染后可获得条带清晰、背景干扰小的图像。该体系的构建为AFLP技术在大菱鲆相关研究中的应用奠定了基础。AFLP （Amplified Fragment Length Polymorphism） analysis system for turbot was established in this study, with the relative processes being presented, including DNA extraction, double enzymes digestion reaction, adapter ligation reaction, pre-amplification and selective amplification reactions, and argent dyeing. It was proved that quite steady results could be achieved by using EcoR I/Mse I double enzymes digestion, three selective base primers, 1 ： 6（ω：ω） EcoR I/Mse I selective primers, and 20 μl volume of selective amplification. The pictures of the production by way of electrophoresis and argent dying were very well, the bands in focus and background less disturbed. The establishment of the system might lay a foundation for the application of AFLP techniques to the relative research of Scophthalmus maximus.

关 键 词：AFLP 大菱鲆 反应体系 

分 类 号：Q7[生物学—分子生物学] Q959.486