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[目的]优化小麦SRAP-PCR技术体系。[方法]以小麦丰优68为试材,利用正交设计L16(45)对小麦SRAP-PCR反应体系中的5因素(Taq聚合酶、Mg2+、模板DNA、dNTPs、引物)在4个水平上进行优化试验。[结果]不同因素对小麦SRAP反应体系影响的大小顺序为:Mg2+>Taq聚合酶>dNTPs>模板DNA>引物;优化的小麦SRAP-PCR体系为:在20μl反应体系中,包括10×PCR buffer2.0μl、Mg2+2.0mmol/L、Taq聚合酶2.0U、dNTPs0.2mmol/L、模板DNA40ng、引物0.6μmol/L。[结论]该优化体系为小麦资源SRAP遗传分析奠定了技术基础。
[Objective] The research aimed to optimize wheat SRAP-PCR technology system. [Method] With Wheat Fengyou 68 as test material, 5 factors (Taq polymerase, Mg2 +, template DNA, dNTPs and primer) in wheat SRAP-PCR system were orthogonal design L16 (45) Optimize the test. [Result] The order of the effect of different factors on SRAP reaction system of wheat was: Mg2 +> Taq polymerase> dNTPs> template DNA> primer. The optimized wheat SRAP-PCR system was: 20 × reaction system including 10 × PCR buffer2. 0 μl, Mg2 + 2.0 mmol / L, Taq polymerase 2.0 U, dNTPs 0.2 mmol / L, template DNA 40 ng, primer 0.6 μmol / L. [Conclusion] The optimized system laid the technical foundation for genetic analysis of SRAP in wheat resources.