甜菜SSR-PCR反应体系的优化  

Optimization of Sugarbeet SSR-PCR Reaction System

作　　者：吴则东[1] 王华忠[1] 韩英[1] 

机构地区：[1]黑龙江省普通高校甜菜遗传育种重点实验室

出　　处：《中国糖料》2008年第1期11-13,17,共4页Sugar Crops of China

基　　金：国家"863"项目(2001AA241192)资助

摘　　要：为了建立适宜甜菜的SSR反应体系,笔者以甜菜不育系TB005A为试验材料,研究了甜菜SSR技术中PCR反应体系的主要成分对SSR扩增结果的影响。对SSR反应体系中的DNA模板浓度、引物浓度、Taq聚合酶浓度、dNTPs浓度以及退火温度进行了探索,确立了适合甜菜的SSR反应体系为:在20μL反应体系中,模板DNA为60ng,引物(50ng/μL)1.2μL,TaqDNA聚合酶1.0U,dNTPs(2.5mM/L)0.6μL。并利用该反应体系对30个甜菜不同品系进行SSR反应,用6%的变性聚丙烯酰胺凝胶电泳检测,不同品系间DNA谱带多态性丰富,证实该体系稳定可靠。In this study, SSR-PCR amplified condition was optimized on the CMS sugarbeet of TB005A and analyzed effects of the concentration of DNA, primer, Taq polymerase, dNTPs and the optimal annealing temperature of primers on PCR results. The results showed that 60ng template DNA, 1.20,L primers （50ng/μL）, 1.0U Taq DNA polymerase and 0.6μL dNTPs （2.5mM/L） in 20μL SSR reaction system were the best suitable PCR system, SSR of 30 different sugarbeet lines were obtained and detected. Polymorphism between different lines abundantly detected by 6% denaturing polyacrylamide gel which showed this system was suitable and stable.

关 键 词：甜菜 SSR SSR-PCR反应条件 变性聚丙烯酰胺凝胶 

分 类 号：S566.301[农业科学—作物学] Q78[生物学—分子生物学]