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以枣树嫩叶为材料,采用单因子试验对影响SRAP-PCR反应体系中的各个因素进行优化,建立可靠、稳定的SRAP-PCR体系,为进一步研究新疆枣种质资源遗传多样性、亲缘关系、指纹图谱的构建等提供帮助。结果表明,最佳SRAP-PCR反应体系为:总体积25.0μL,其中模板DNA用量为30 ng,Mg2+浓度为2.5 mmol/L,引物浓度为0.2μmol/L,d NTPs浓度为0.2 mmol/L,Taq DNA聚合酶用量为1.0 U,该体系经验证,能扩增出清晰、稳定、多态性较好的产物,说明经过优化后的PCR体系较好,适合后续的分析研究。
In order to further study the genetic diversity and genetic relationship of jujube germplasm resources in Xinjiang, single-factor experiments were used to optimize the factors that affect the SRAP-PCR reaction system and to establish a reliable and stable SRAP-PCR system. , Fingerprinting and other structures to provide help. The results showed that the best SRAP-PCR reaction system was as follows: total volume 25.0μL, template DNA 30 ng, Mg2 + 2.5 mmol / L, primer 0.2μmol / L, d NTPs 0.2 mmol / The amount of Taq DNA polymerase was 1.0 U. The system was verified to amplify a clear, stable and better polymorphic product, indicating that the optimized PCR system is better suited for subsequent analysis.