通江百合SRAP-PCR体系优化及引物筛选  

Optimization of SRAP-PCR System and Selection of Primers for Lilium sargentiae

作　　者：智丽[1] 滕中华[2] 李先源[1] 眭顺照[1] 李名扬[1] 

机构地区：[1]西南大学园艺园林学院,重庆市花卉工程技术研究中心,南方山地园艺学教育部重点实验室,重庆400716 [2]西南大学农学与生物科技学院,重庆400716

出　　处：《西南大学学报（自然科学版）》2013年第4期32-38,共7页Journal of Southwest University(Natural Science Edition)

基　　金：国家自然科学基金(30872063);中央高校基本科研业务费专项资金(XDJK2010C077);西南大学博士启动基金项目(swu112023)资助

摘　　要：利用正交设计和单因素试验相结合的方法,从模板DNA,引物,dNTPs,Taq DNA聚合酶,Mg2+等5因素4水平来优化通江百合SRAP-PCR反应体系,优化后的反应体系为Mg2+2.5mmol/L,Taq DNA聚合酶0.25U,dNTPs 0.25μmol/L,引物0.25μmol/L,模板DNA 50ng,总体积为25μL.依据优化后的体系从144对引物中筛选出扩增条带清晰,多态性丰富的引物组合24对,体系验证结果表明,优化后的体系具有较高的稳定性和重复性.Orthogonal experimental design and single factor experiment were applied in combination to study the effects of 4 levels of DNA template, primers, dNTPs, Taq DNA polymerase and Mg2＋ on SRAP-PCR for Lilium sargentiae. The concentration of Mg2＋ was shown to be the most dominant factor for the results of SRAP PCR, while the concentration of DNA template from 20 ng to 50 ng had almost no effects on it. An optimized SRAP-PCR system for L. sargentiae was established, which contained 2.5 mmol/L Mg2＋ 0. 25 U Taq DNA polymerase, 0.25 μmol/L dNTPs , 0.25 μmol/L primers and 50 ng DNA template in a total volume of 25μL. Based on the optimized system, 24 primer combinations were selected from 144 primer pairs, which had abundant polymorphism bands, and the annealing temperature of the selected primer combinations was also screened. The system was proved to be stable and repeatable with the different samples of L. sargentiae derived from various regions.

关 键 词：通江百合 体系优化 SRAP 

分 类 号：Q949.718.23[生物学—植物学] S682.29[农业科学—观赏园艺]