花生SSR-PCR体系的优化  

Optimization of SSR – PCR Reaction System of Peanut

作　　者：张发[1] 万勇善[1] 刘风珍[1] 

机构地区：[1]山东农业大学农学院作物生物学国家重点实验室,泰安271018

出　　处：《中国农学通报》2008年第4期37-41,共5页Chinese Agricultural Science Bulletin

基　　金：国家863计划项目(编号:2006AA10A115);山东省中青年科学家科研奖励基金项目(编号:2006BSB01049)

摘　　要：以花生品种丰花3号为材料,研究了花生SSR技术中PCR反应体系的主要成分对SSR扩增效果的影响及不同引物的退火温度。结果表明dNTP对扩增影响较大,每对引物都有其扩增适合的退火温度。确立了适合花生SSR分子标记研究的优化体系。最终确定总反应体系为20μl,其中25mMMgCl21.5μl,2.5mMdNTP1.6μl,5U/μlTaq酶0.17μl,100ng/μl模板DNA0.4μl,2.5mM引物5μl。优化后的扩增程序退火温度为54℃。FengHua 3 was used in this study to optimize the several factors applied in SSR technique system. dNTP was found to have distinct effect to amplification, and annealing temperature specific to primer pair was also checked. Optimized SSR system for peanut was established ： 25 mM MgCl2 1.5 μl, 2.5 mM dNTP 1.6 μl, 5 U/μl Taq 0.17 μl, 100 ng/μl template DNA 0.4 μl, 2.5 mΜ primers 5 μl were added to the PCR mixture. Optimized annealing temperature was 54℃.

关 键 词：花生 SSR 体系优化 

分 类 号：S565.2[农业科学—作物学] Q341[生物学—遗传学]