摘要
以广西地方桑树种质资源"平武1号"为材料,以引物组合Me8/Em3对其SRAP反应体系的模板DNA、引物、dNTPs和Taq DNA聚合酶等条件进行优化。结果表明,最优反应体系为:在25.0μL反应体系中含模板DNA 45.0 ng,引物1.0μmol/L,dNTPs 0.3 mmol/L,Taq酶1.0 U。用42个引物组合对6份桑树二倍体品系(农桑8号、璜桑37号、湖桑197号、湖桑32号、湖桑199号、育711、桐乡青、国桑20号)及其同源四倍体进行扩增,其中引物组合Me6/Em2的扩增条带清晰,品种间有不同程度的多态性。该研究建立的反应体系应用于桑树遗传多样性研究,重复性好,稳定性强,结果可靠,可用于下一步研究。
The experiment was conducted to optimize the concentration of template DNA,primer,dNTPs and Taq DNA polymerase in SRAP reaction system for mulberry variety Pingwu 1.The results showed that the optimum reaction system in a 25.0 μL PCR mixture consisted oaf 45 ng genomic DNA,1.0 μmol/L primers,0.3 mmol/L of dNTPs,and 1 U Taq polymerase.Fourty-two primer combinations were used to conduct the SRAP-PCR amplification of six mulberry diploid species and their autotetraploids.The results indicated clear amplified bands showing polymorphism among different species when amplified with primer Me6/Em2.The PCR method developed in this research showed good repeatability and high stability when applied to study the genetic diversity in mulberry,and could be used in the future research.
出处
《广西农业科学》
CSCD
2010年第11期1151-1154,共4页
Guangxi Agricultural Sciences
基金
广西自然科学基金项目(桂科自0991177)
