摘要
采用改良的CTAB法提取林下参的基因组DNA,所得的DNA纯度高、质量好,可用于RAPD分析.筛选出的林下参RAPD反应的最佳体系为20μL,反应体系中包括模板DNA 20ng,引物20 pmol,dNTPs 0.187 5 mmol/L,TaqDNA聚合酶1.5 U,Mg2+2.0 mmol/L,10×Reaction Buffer 2.0 mmol/L,其余部分用无菌超纯水补充.PCR扩增程序为94℃预变性5min,94℃变性1 min,37℃退火1 min,72℃延伸2 min,40次循环,72℃最终延伸7 min.应用优化后的反应体系PCR扩增获得的RAPD指纹图谱带型清晰,重复性好,为通过分子标记获得丰富的林下参遗传信息奠定了基础.
High purity and quality DNA extracted of Transplanted wild ginseng was used for RAPD analysis. The best RAPD reaction system optimized was： total volume 20 μL, containing template DNA 20 ng, primer 20 pmol, dNTPs 0. 187 5 mmol/L, MgCI2 2.0 mmol/L, DNA polymerase 1. 5 unit, 10× Reaction Buffer 2.0 mmol/L, completed with ddH20. PCR amplification condition was： pre-denaturation at 94 ℃for 5 min, denaturation at 94 ℃ for 1 min, annealing at 37℃ for 1 rain, extension at 72 ℃ for 2 min, cycle for 40 times. Final extension at 72℃ for 7 rain. RAPD finger printing obtained by the optimized system was clear and repetitive, which provided a certain foundation for getting more extensive genetic information of Transplanted wild ginseng of Changbai Mountain.
出处
《延边大学农学学报》
2009年第1期16-20,52,共6页
Agricultural Science Journal of Yanbian University
基金
延边州科技发展基金资助项目(2008YB501A16)
