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

小麦SRAP-PCR体系的正交设计优化(摘要)(英文)

[Objective] The aim was to optimize the technical procedure of SRAP-PCR in Triticum aestivum L. [Method] The orthogonal design L16（45） 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 polymerasedNTPsDNA templateprimer. 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 Taq polymerase,2.0 mmol/L Mg2＋,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 conduct SRAP genetic analysis for wheat varieties.

[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。