黄精SRAP-PCR反应体系的优化及引物筛选  

Optimization of SRAP-PCR System for Polygonati Rhizoma and Screening of Effective Primers

作　　者：陈卫易 徐昌艳[1] 钱志瑶 吴红 王友峰 覃容贵[1] 罗忠圣 CHEN Weiyi;XU Changyan;QIAN Zhiyao;WU Hong;WANG Youfeng;QIN Ronggui;LUO Zhongsheng(Guizhou Medical University,Guiyang 550025,China;Natural Products Research Center of Guizhou Province,Guiyang 550014,China)

机构地区：[1]贵州医科大学,贵州贵阳550025 [2]贵州省天然产物研究中心,贵州贵阳550014

出　　处：《山东化工》2025年第6期4-8,共5页Shandong Chemical Industry

基　　金：贵州省大学生创新训练项目计划(编号:S202210660141)。

摘　　要：目的:优化并建立黄精SRAP反应体系,为黄精遗传多样性研究及其资源保护、良种选育提供更多理论依据。方法:在单因素试验的基础上,以Taq PCR MasterMix用量、Mg^(2+)浓度、模板DNA用量、Taq DNA聚合酶用量进行正交试验设计,优化黄精SRAP-PCR反应体系,并进行有效引物筛选。结果:黄精的SRAP最佳反应体系为:12μL Taq PCR Master Mix,30 ng模板DNA,1.25μmol/L引物,0.375 mmol/L Mg^(2+),0.17 mmol/L dNTPs,1.5 U Taq DNA聚合酶,加入ddH_(2)O至20μL;从99对随机引物中筛选出7对有效引物。结论:建立的黄精SRAP-PCR反应体系稳定性、重复性良好,可用于黄精遗传多样性研究。Objective:To establish the SRAP reaction system and study the genetic diversity of Polygonati rhizoma,so as to lay the theoretical and technical foundations for the conservation of its resources and and the selection and breeding of its varieties.Methods:On the basis of single factor test,the orthogonal assay design was performed to optimize the effects of Taq PCR MasterMix dose,Mg^(2+)concentration,template DNA concentration,and Taq DNA polymerase content on SRAP-PCR amplification of Polygonati rhizoma.And performed an effective primer screening.Results:The optimal SRAP-PCR reaction system for Polygonati rhizoma was:12μL Taq PCR Master Mix,30 ng of template DNA,1.25μmol/L primer,0.375 mmol/L Mg^(2+),0.17 mmol/L dNTPs,1.5 U Taq DNA polymerase,and ddH_(2)O was then added to a final total volume of 20μL.Out of 99 pairs of SRAP-PCR primers,7 pairs of effective primers were available for genetic diversity analysis of Polygonati rhizoma.Conclusion:The SRAP-PCR reaction system of Polygonati rhizoma has good stability and reproducible,which can be used for the study of genetic diversity.

关 键 词：黄精 相关序列扩增多态性 分子标记 遗传多样性 

分 类 号：TQ461[化学工程—制药化工]