摘要
[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反应体系的影响为: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×PCRBuffer2.0μl、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 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, Mg^2, DNA template, dNTPs and primer） at four levels. [ Result] Effects of the five factors on SRAP-PCR reaction system were Mg^2 〉 Taq polymerase 〉 dNTPs 〉 DNA template 〉 primer. Finally, an optimal SRAP-PCR system was established, that was the total 20μl reaction system containing 2.0μ1 10×PCR Buffer, 2.0 U DNA polymerase, 0.2 mmol/L dNTPs, 40 ng DNA template and 0.6μmol/L primer. [ Conclusion] The optimum SRAP-PCR system had provided some technical foundations to study the phylogenetic relationship of wheat varieties.
出处
《安徽农业科学》
CAS
北大核心
2011年第17期10121-10123,共3页
Journal of Anhui Agricultural Sciences
基金
国家科技支撑计划项目(2008FY110500-9)
