| 大竹蛏AFLP分子标记反应体系的建立与优化 |
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| 引用本文： | 张滔,刘相全,孙国华,刘丽娟,孙振兴.大竹蛏AFLP分子标记反应体系的建立与优化[J].安徽农业科学,2011,39(29):17923-17926. |
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| 作者姓名： | 张滔 刘相全 孙国华 刘丽娟 孙振兴 |
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| 作者单位： | 鲁东大学生命科学学院,山东烟台,264025;山东省海洋水产研究所,山东烟台,264006 |
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| 基金项目： | 山东省农业良种工程项目“优质高产抗逆贝类良种选育” |
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| 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。
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| 关 键 词： | 大竹蛏 AFLP 分子标记 |
The Establishment and Optimization of AFLP Molecular Marker Reaction System for Solen grandis |
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| Institution: | ZHANG Tao et al(College of Life Science,Ludong University,Yantai,Shandong 264025) |
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| Abstract: | 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. |
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| Keywords: | Solen grandis AFLP Molecular marker |
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