摘要
以葡萄新鲜幼嫩叶片为材料,研究了SSR分析过程中的影响因素,包括Taq酶、Mg^2+、dNTP、引物、模板DNA浓度、退火温度等,建立了适合葡萄SSR反应的PCR体系,即20m反应体系中含有Taq酶1.0U、Mg^2+浓度为1.5mmol/L,dNTP0.2mmol/L,引物0.5/μmol/L,模板DNA20—30ng。扩增程序为:95℃预变性4min;95℃变性50s、50~60℃退火1 min、72%延伸1 min30s,30个循环;最后72℃延伸7min。
The factors influencing SSR analysis, including Taq polymerase, Mg^2+ , dNTP, primers, template DNA' s concentration, annealing temperature and so on were studied using fresh young leaves of grape. An optimal PCR system for SSR in fresh leaves of grape has been found:in 20μL reaction solution, contained 1.0 U Taqpolymerase, 1.5 mmol/L of Mg^2+ ,0. 2 mmol/L of dNTP,0.5 μmol/L of primers, and 20 - 30 ng of template DNA. The amplification program was devised:initial denaturing at 95℃ for 4min; denaturing at 95℃ for 50 s, annealing at 50℃ - 60℃ for 1 min, and extension at 72℃ for 1 min 30 s,30 cycles;final extension at 72℃ for 7 min.
出处
《河北林果研究》
2008年第3期281-286,共6页
Hebei Journal of Forestry and Orchard Research
基金
河北省林业局项目
关键词
葡萄
SSR
优化
grape
SSR
optimization