扁玉螺ISSR-PCR反应体系的建立及其优化  

Establishment and Optimization of the ISSR-PCR Reaction System for Neverita didyma

作　　者：孙始威[1] 孙振兴[1] 陈燕妮[1] 吴克 

机构地区：[1]鲁东大学生命科学学院,山东烟台264025 [2]烟台经济技术开发区第八小学,山东烟台264006

出　　处：《湖北农业科学》2008年第5期575-577,共3页Hubei Agricultural Sciences

摘　　要：以扁玉螺基因组DNA为材料,分析了Mg2+、dNTP、Taq DNA聚合酶、引物以及模板DNA浓度对ISSR-PCR扩增效果的影响,建立了一套适合扁玉螺的ISSR-PCR反应体系。该反应体系包括:2.0mmol·L-1的Mg2+,0.25 mmol·L-1的dNTP,0.8 U的Taq DNA聚合酶,0.2μmol·L-1的ISSR引物和40 ng模板DNA。利用优化后的反应体系,从30条ISSR引物中筛选出13条扩增效果较好的引物,为进一步利用ISSR标记研究扁玉螺的遗传多样性奠定了基础。The influence of Mg^2＋, dNTP, Taq DNA polymerase, ISSR primer and template DNA concentration on the result of ISSR-PCR amplification was analyzed using the genomic DNA of Neverita didyma. The ISSR-PCR reaction system was established for Neverita didyma. The reaction system was as follows： 2.0 mmol·L^-1 Mg^2＋, 0.25 mmol·L^-1 dNTP, 0.8 U Taq DNA polymerase, 0.2μmmol·L^-1 ISSR primer, 40 ng template DNA. Using the optimized reaction system, 13 primers with good polymorphism screened from 30 ISSR primers were used to conduct ISSR analysis. The result provided basis for further the analysis of genetic diversity of Neverita didyma by ISSR marker.

关 键 词：扁玉螺 ISSR—PCR 引物筛选 

分 类 号：Q75[生物学—分子生物学] S961.6[农业科学—水产养殖]