小麦SRAP-PCR体系的正交设计优化

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（4^5）对小麦SRAP．PCR反应体系中的5因素（细聚合酶、Mg^2、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反应体系的影响为：Mg^2〉Taq聚合酶〉dNTPs〉模板DNA〉引物；优化的小麦SRAP—PCR体系为：在20山反应体系中，包括10×PCRBuffer2．0山、Mg^2 2．0mmol／L、砌聚合酶2．0U、dNTPs0．2mmol／L、模板DNA40ng、引物0．6μmolVL。结论]该优化体系为小麦资源SRAP遗传分析奠定了技术基础。

Optimization of SRAP-PCR System of Triticum aestivum-Based on Orthogonal Design

Objective] The aim was to optimize the technical procedure of SRAP-PCR in Triticum aestivum L.. Method] The orthogonal design was used to optimize SRAP-PCR amplification system of wheat Fengyou 68 on five factors (Taq polymerase, Mg2+, DNA template, dNTPs and primer) at four levels. Result] Effects of the five factors on SRAP-PCR reaction system were Mg2+ >Taq polymerase>dNTPs> DNA template > primer. Finally, an optimal SRAP-PCR system was established, that was the total 20 μl reaction system containing 2.0 μl 10×PCR Buffer, 2.0 U DNA polymerase, 0.2 mmol/L dNTPs, 40 ng DNA template and 0.6 μmol/L primer. Conclusion] The optimum SRAP-PCR system had provided some technical foundations to study the phylogenetic relationship of wheat varieties.