澳洲坚果SSR-PCR反应体系优化及其应用

Optimization and Application of SSR-PCR Reaction System for Macadamia

摘要: 以澳洲坚果DNA为模板，通过单因素设计方法对影响SSR-PCR反应体系的5个主要因素进行了优化。结果表明，澳洲坚果SSR-PCR反应体系的最适条件为：20 μL的反应体系中，包含30 mg·L^-1模板DNA，1.0 U Taq聚合酶，2.5 mmol·L^-1 Mg²⁺，0.6 μmol·L^-1引物和0.3 mmol·L^-1dNTPs。采用该反应体系对不同引物和15份澳洲坚果种质进行验证，扩增条带的可靠性和稳定性良好，且分辨率较高。因此，该SSR-PCR反应体系可用于澳洲坚果种质源鉴定及遗传多样性分析研究。

Abstract: Five major factors of the SSR-PCR reaction system were optimized for genomic DNA of macadamia (Macadamia spp.) by a single factor design. The volume of optimum reaction system was 20 μL that consisted of 30 mg·L^-1 template DNA, 1.0 U Taq polymerase, 2.5 mmol·L^-1 Mg²⁺, 0.6 μmol·L^-1 primer and 0.3 mmol·L^-1 dNTPs. On 15 germplasms of macadamia using different SSR primers, the system proved to be reliable and stable in the amplification bands with high resolution. Consequently, it seemed adequate for the identification and genetic diversity analysis of macadamia germplasms.