摘要
本研究先采用单因素试验确定5个影响因素的浓度区域,然后进一步进行正交试验筛选各因素最优水平,通过两种实验方法相结合,优化、建立流苏树属(Chionanthus)SSR-PCR反应体系,最后检测其稳定性。研究结果表明,优化后的20μL流苏树属SSR-PCR反应体系为:10×Buffer 2μL,DNA模板2.0 ng/μL,Mg2+2.5 mmol/L,Taq酶2.5 U,dNTPs 250μmol/L,引物0.3μmol/L(正反引物各0.15μmol/L),ddH2O补足。用4份材料对反应体系进行验证均可得到多态性丰富、清晰稳定的条带,证明该反应条件可用于流苏树属的遗传多样性分析,可为流苏树属的遗传多样性研究、遗传变异分析、亲缘关系探讨及种质资源的科学保护奠定坚实的基础提供基础数据。
In this study,the single factor test was used to determine the concentration area of five factors,and then further performing the orthogonal test to screen the optimal level of each factor.The SSR-PCR reaction system was established by combining the two experimental methods,and the stability of the SSR-PCR reaction system was finally detected.The results showed that the optimized 20μL SSR-PCR reaction system included 10×Buffer2μL,DNA template 2.0 ng/μL Mg2+2.5 mmol/L,Taq DNA polymerase 2.5 U,dNTPs 250μmol/L,Primer 0.3μmol/L(the forward primer and reverse primer were 0.15μmol/L each),and added ddH2 O to 20μL at last.The four materials were used to verify the reaction system,and the bands with rich polymorphism and clear and stable were obtained.This results showed that the reaction conditions could be used to analyze the genetic diversity of the genus tassel,which could provid basis data for genetic diversity research,genetic variation analysis,relationship discussion and scientific protection of germplasm resources of Chionanthus.
作者
李晓
张鸽香
Li Xiao;Zhang Gexiang(College of Landscape Architecture,Nanjing Forestry University,Nanjing,210037)
出处
《基因组学与应用生物学》
CAS
CSCD
北大核心
2020年第11期5247-5254,共8页
Genomics and Applied Biology
基金
国家林业局948项目美国流苏优良种质繁育技术引进(2014-4-17)
江苏高校品牌专业建设工程资助项目(PPZY2015A063)共同资助。
