大竹蛏AFLP分子标记反应体系的建立与优化  

The Establishment and Optimization of AFLP Molecular Marker Reaction System for Solen grandis

作　　者：张滔[1] 刘相全[2] 孙国华[2] 刘丽娟[2] 孙振兴[1] 

机构地区：[1]鲁东大学生命科学学院,山东烟台264025 [2]山东省海洋水产研究所,山东烟台264006

出　　处：《安徽农业科学》2011年第29期17923-17926,共4页Journal of Anhui Agricultural Sciences

基　　金：山东省农业良种工程项目"优质高产抗逆贝类良种选育"

摘　　要：[目的]建立一套适合大竹蛏的AFLP分子标记反应体系,为大竹蛏的遗传多样性研究提供技术支持。[方法]采用比较实验法,以大竹蛏基因组DNA为模板,对其AFLP体系中的酶切时间、预扩产物稀释倍数及选扩引物E+3/M+3的配比等进行优化。[结果]优化的AFLP体系为:酶切时的DNA模板浓度为100 ng/μl,双酶切2 h;预扩模板最佳用量1.0μl;预扩产物的最适稀释倍数为40倍;选扩引物浓度配比为1∶4。同时,筛选出12对引物组合,可用于大竹蛏的AFLP分析。[结论]采用优化的AFLP反应体系及筛选后的引物,对大竹蛏群体进行了初步扩增,得到了清晰的电泳图谱。[Objective] The research aimed to establish the operation rules for AFLP-PCR reaction system in Solen grandis and provide basis for its application in genetic diversity.[Method] With comparing experimentation,by using genomic DNA of Solen grandis adjusting the time for DNA double enzymes digestion reaction,the dilute multiple for the per-amplification production and different proportion in selective primers M＋3/E＋3 to select the optimization.[Result] The optimum AFLP system was as followed： concentration of template DNA is 100 ng /μl in the digest system and 2 hours for the reaction;1.0 microlitre ligation products as the template of pre-amplification;40 times of dilution for the products of pre-amplification with 1×TE for selective amplification;the ratio of primer concentration was 1∶4.And 12 pairs of informative primers were identified and recommended for AFLP analysis of Solen grandis.[Conclusion] The PCR amplification of Solen grandis were analyzed by using the optimized AFLP reaction system and most primers amplified clear bands.

关 键 词：大竹蛏 AFLP 分子标记 

分 类 号：Q959.21[生物学—动物学]