高粱SCoT-PCR反应体系的建立与优化

Establishment and Optimization of SCoT-PCR Reaction System for Sorghum

为利用分子标记辅助育种技术更好地保护高粱的种质资源,本试验构建了高粱SCoT-PCR的最佳反应体系。试验采用单因素实验设计的方法对影响高粱SCoT-PCR反应体系的5个因素,即dNTP用量、DNA的用量、Mg2+浓度、引物用量、Taq DNA聚合酶浓度分别进行梯度设计优化分析,并对得到的最佳体系进行验证。综合单因素试验结果,明确在高粱SCoT-PCR 25μL反应体系中,最佳反应体系为:引物2.0μL,DNA 1.25μL,dNTP 1.0μL,Taq DNA聚合酶0.2μL,Mg2+2.0μL。本试验研究结果可为高粱的遗传多样性研究提供良好研究基础,并为高粱种质保护、育种提供帮助。

In order to use molecular marker-assisted breeding technology to better protect the germplasm resources of sorghum,this experiment constructed the best reaction system of sorghum SCoT-PCR.The single-factor experimental design method is used to influence the 5 factors of sorghum SCoT-PCR reaction system,namely dNTP dosage,DNA dosage,Mg2+concentration,primer dosage,and Taq DNA polymerase concentration were respectively performed gradient design optimization analysis,and the best system obtained was verified.Based on the single factor test results,it is clear that in the 25μL reaction system of sorghum SCoT-PCR,the best reaction system is:primer 2.0μL,DNA 1.25μL,dNTP 1.0μL,Taq DNA polymerase 0.2μL,Mg2+2.0μL.The research results can provide help for future development of sorghum germplasm resources protection and molecular marker assisted breeding.The results of this experiment can provide a good research foundation for the genetic diversity research of sorghum,and provide help for the protection and breeding of sorghum germplasm.