基因组DNA混合池技术在SSR引物筛选中的可靠性分析

Reliability Analysis of Genomic DNA Mixing Pool Technology in SSR Primer Screening

SSR-PCR产物的重测序是SSR标记技术开发和应用的必要步骤。本研究以6个桉树DNA样品为对照组,1个基因组DNA混合池为试验组进行基因组DNA混合池在SSR引物筛选中的可靠性性分析。通过SSR-PCR产物的重测序分析,6个桉树单一DNA样品与基因组DNA混合池的扩增序列比对结果分析显示,两种模板扩增结果差异不显著,即基因组DNA混合池可以替代多个单一DNA样品进行SSR引物的高效筛选。利用20对SSR引物在6种桉树DNA样品中的扩增序列进行多态性分析发现,同一位点不同桉树种间的扩增序列存在长度多态性和核苷酸多态性,从而证实了SSR序列的多态性和复杂性,为SSR标记技术在桉树遗传育种研究中的应用提供了理论依据。

The S SR-PCR products sequencing is an essential step in development and application of SSR markers. In the research, 6 Eucalyptus DNA samples were used as the control group, and a genomic DNA mixing pool was selected as the treatment group to analyze the reliability of genomic DNA pooling in SSR primers screening. Through the sequencing alignment analysis of SSR-PCR products, analysis of amplification sequence of 6 Eucalyptus single DNA samples and genomic DNA mixing pool showed that the amplification of two templates had no significant difference. That was to say, the genomic DNA mixing pool could be used for efficient screening of SSR primers instead of multiple single DNA samples. Using 20 pairs of SSR primers, the amplified sequences of 6 Eucalyptus DNA samples were analyzed by polymorphism. It was found that there were polymorphism and nucleotide polymorphism in the amplified sequences among different Eucalyptus species at the same site. Thus, the polymorphism and complexity of SSR sequences were confirmed, which provided a theoretical basis for the application of SSR markers in the study of Eucalyptus genetic breeding.