ISSR标记在扁玉螺中的初步应用

Preliminary application of ISSR technique in Neverita didyma

以扁玉螺基因组DNA为材料,分析了Mg2+、dNTP、TaqDNA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的Mg2+,0.25mmol/L的dNTP,0.8U的TaqDNA聚合酶,0.2μmol/L的ISSR引物和40ng模板DNA。利用优化后的反应体系,从30条ISSR引物中筛选出了13条扩增效果较好的引物,这为进一步利用ISSR标记研究扁玉螺的遗传多样性打下了基础。

The influence of Mg^2＋, dNTP, Taq DNA polvmerase, 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 m mol/L Mg^2＋, 0.25 m mol/L dNTP, 0.8 U Taq DNA polymerase, 0.2 μmol/ L ISSR primer, 40 ng template DNA. Using the optimized reaction system, 13 primers with Rood polvmorphism 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.