叶子花SSR-PCR体系的优化  

Optimization of SSR-PCR System for Bougainvillea arborea

作　　者：鲁亚静 陈庭[3] 李建友[3] 胡章立[1] 昝启杰[1,2] 陈涛[1,3] 

机构地区：[1]深圳大学生命科学学院,广东深圳518060 [2]深圳市野生动物救护中心,广东深圳518040 [3]深圳市中国科学院仙湖植物园,广东深圳518004

出　　处：《安徽农业科学》2015年第17期47-49,53,共4页Journal of Anhui Agricultural Sciences

基　　金：深圳市科技研发资金项目(JCYJ20120615172425764);深圳市城管局科研项目(200901)

摘　　要：[目的]优化叶子花SSR-PCR体系。[方法]以乔木叶子花为材料,通过Touchdown PCR扩增程序和正交试验,对影响叶子花SSR-PCR体系的引物浓度、Taq DNA聚合酶用量、Mg2+浓度和d NTPs浓度进行优化。[结果]叶子花20μl SSR-PCR体系的最优条件为引物浓度0.4 mmol/L、Taq DNA聚合酶用量1.5 U、Mg2+浓度1.5 mmol/L、4种d NTPs浓度均为0.15 mmol/L,模板100 ng左右。[结论]该反应体系的扩增条带清晰、稳定、重复性好,可用于后续叶子花遗传多样性分析和种质资源鉴定等研究。[ Objective ] The aim was to optimize SSR-PCR system for Bougainvillea arborea by orthogonal design. [ Method ] Based on orthogo- nal design, four levels of four factors （primer, Taq DNA polymerase, Mg^2＋ , dNTPs） have been tested to optimize SSR-PCR reaction system using Bougainvillea aborea genomic DNA as template. The amplification was performed with a touchdown program. [ Result ] The optimized SSR-PCR reaction condition for Bougainvillea arborea was obtained in a 20 μl system containing 0.4 mmol/L SSR primer, 1.5 U Tat/DNA polymerase, 1.5 mmol/L Mg^2＋ , 0. 15 mmol/L dNTPs and around 100 ng DNA template. [ Conclusion] With clear, stable and repeatable am- plification bands, the reaction system can be used for the further studies on genetic diversity and germplasm identification.

关 键 词：叶子花 正交试验 SSR PCR体系优化 

分 类 号：S188[农业科学—农业基础科学]