摘要
采用正交设计法对影响美花石斛SSR-PCR体系的5个主要因素进行了优化,建立适宜的美花石斛SSR-PCR的反应体系,并利用黔产5个不同的美花石斛居群共计48株进行了验证。结果表明:在10μL的美花石斛的反应体系中各因素的最佳浓度分别为2.0mmol/L Mg^(2+),0.4mmol/L dNTP,1.5UTaq酶,0.4μmol/L引物,40ng模板DNA,建立的SSR-PCR体系可用于美花石斛的遗传多样性研究。
In order to establish the best SSR-PCR reaction system, by the method of the orthogonal design affect on D. loddigesii SSR-PCR system of 5 elements(Mg2+ , dNTP, Taq enzyme, primers and DNA) was optimized,which estabiished suitable D.loddigesii SSR-PCR reaction system.And a total of 48 species of D. loddigesii Rolfe from 5 group in guizhou were tested.The results showed the optimum concentration of each factor in the reaction system of D.loddigesii in I0 pL volumes was 2.0 mmol/L Mg2+ ,0.4 mmol/L dNTP, 1.5 U Taq DNA polymerase,0.4μmol/L primer and 40 ng template DNA.The established SSR-PCR system can be used for analyzing genetic diversity of D,loddigesii.
作者
张文龙
石春杰
ZHANG Wenlong;SHI Chunjie(Pharmacy Department, Guiyang College of Traditional Chinese Medicine, Guiyang Guizhou 550025, China;Guizhou Institute of Upland Food Crops,Guizhou Academy of Agricultural Sciences, Guiyang 550006, China)
出处
《种子》
北大核心
2018年第4期23-25,共3页
Seed
基金
贵州省科技基金(黔科合J字[2013]2072号)
黔教合人才团队字[2013]12号
贵阳中医学院博士资金项目(贵中医博[2012]01号)
关键词
美花石斛
SSR正交设计
D.loddigesii Rolfe
SSR
orthogonat design