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.
出处
《湖北农业科学》
北大核心
2008年第5期575-577,共3页
Hubei Agricultural Sciences
