摘要
以阿尔冈金等6个地方主栽的紫花苜蓿品种为材料,对随机扩增多态性DNA标记(random amplified polymorphic DNA,RAPD)法鉴定不同紫花苜蓿品种的试验条件进行了优化。结果表明,使用改良的CTAB法提取紫花苜蓿基因组DNA蛋白较少,质量较高;PCR扩增反应体系中使用1 U/μL的TaqDNA聚合酶、10 ng/μL的DNA模板和300μmol/L的dNTP,扩增的条带较多,且较清晰,适宜于后续RAPD法鉴定;从100条10个碱基的随机引物中筛选出S37与S140两条引物,适用于6个不同苜蓿品种的鉴定。
In this study, RAPD（Random Amplified Polymorphic DNA） optimal parameters for identifying various alfalfa varieties including the extract of genomic DNA, concentration of Taq DNA polymerase, DNA template and dNTP for PCR amplification were screened. The results showed that the improved CTAB approach, 1 U/μL Taq DNA polymerase, 10 ng/μL DNA template and 300 μmol/L dNTP were pinpointed as a combination of optimal parameters for the RAPD identification of various alfalfa varieties. Subsequently, One hundred 10-mer random primers were used for this screening to distinguish various alfalfa varieties based on the optimal parameters, and the two primers, S37 and S140, were obtained to achieve the goal. The study suggested that this method would provide a new alternative strategy for identifying and breeding alfalfa varieties.
出处
《湖南农业科学》
2016年第1期1-5,共5页
Hunan Agricultural Sciences
基金
国家自然科学基金资助项目(31260568
31160060)
天水市科技局2010年科技支撑计划项目
