苦豆子RAPD反应体系的优化  

Optimization of RAPD reaction system in Sophora alopecuroides L.

作　　者：杨玉蓉[1] 罗慧彬[1] 马渊[1] 刘萍[1] 

机构地区：[1]宁夏大学农学院,宁夏银川750021

出　　处：《农业科学研究》2011年第2期26-29,共4页Journal of Agricultural Sciences

基　　金：国家自然科学基金资助项目(30660017);2009年宁夏大学国家级大学生创新研究项目(091074903)

摘　　要：以苦豆子基因组DNA为模板,采用单因素递进筛选和全面正交设计相结合的方法,获得苦豆子RAPD扩增反应的优化体系.结果表明:在20μL反应体系中,DNA模板20 ng,引物浓度为30μmol/L,Mg2+浓度为50μmol/L,Taq DNA聚合酶2.5 U/μL,dNTP浓度为5 mmol/L,10×PCR Buffer 2.0μL.优化后的体系扩增产物稳定,条带清晰.利用随机引物09M27412对5份苦豆子DNA进行扩增,证实该体系重复性好,结果稳定,可作为苦豆子遗传多样性分析的1个稳定体系.Combining with the method of single factor one by one screening and entire orthogonal design screening,the RAPD-PCR optimum reaction system of Sophora alopecuroides was obtained.The results showed that the optimum concentration of five important components i.e.MgCl2,primer,dNTPs,Taq DNA polymerase,and template DNA in 20 μL reaction system were 50 μmol/L,30μmol/L,5mmol/L,2.5U/μL,and 20ng respectively.There were stable and clear amplified bands in optimum system.Meanwhile,5 genomic DNAs from S.alopecuroides were amplified by using this system with random primers 09M27412,which confirmed this to be stable and have a high reproducibility.It can be utilized in the study of genetic diversity on S.alopecuoides.

关 键 词：苦豆子 RAPD 正交设计 体系优化 

分 类 号：Q943.2[生物学—植物学]