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人参SRAP-PCR体系优化条件的建立
引用本文:许永华,王士杰,陈晓林,史俊卿,杜跃中,张连学. 人参SRAP-PCR体系优化条件的建立[J]. 安徽农业科学, 2010, 38(16): 8419-8420,8439. DOI: 10.3969/j.issn.0517-6611.2010.16.050
作者姓名:许永华  王士杰  陈晓林  史俊卿  杜跃中  张连学
作者单位:吉林农业大学中药材学院,吉林长春,130118;吉林人参研究院,吉林通化,134001
基金项目:国家自然科学基金项目 
摘    要:[目的]研究人参基因组SRAP-PCR的扩增条件,建立其优化扩增体系。[方法]采用单因子实验方法探讨模板DNA、引物浓度、dNTPs浓度、Mg^2+浓度等因素对PCR结果的影响。[结果]优化后的扩增程序为:94℃预变性2 min;94℃变性30 s,48℃复性30 s,72℃延伸1 min,共40次循环;72℃延伸7 min。最佳反应体系为:DNA模板30 ng,上下游引物浓度2.0μmol/L,dNTPs浓度0.3 mmol/L,Mg^2+2.5 mmol/L,总体积25μl。[结论]建立了满足人参SRAP-PCR的优化扩增体系,为人参亲缘关系和遗传多样性SRAP分析提供快速、简便、重复性好的实验方法。

关 键 词:SRAP  PCR  人参

Establishment of the Optimization Conditions of SRAP-PCR System in Ginseng
XU Yong-hua. Establishment of the Optimization Conditions of SRAP-PCR System in Ginseng[J]. Journal of Anhui Agricultural Sciences, 2010, 38(16): 8419-8420,8439. DOI: 10.3969/j.issn.0517-6611.2010.16.050
Authors:XU Yong-hua
Affiliation:XU Yong-hua(College of Chinese Medicine Materials,Jilin Agricultural University,Changchun,Jilin 130118)
Abstract:[Objective] The research aimed to study the amplification conditions of SRAP-PCR of ginseng genome and set up the optimized amplification system.[Method] The effects of template DNA,primer,dNTPs and Mg2+ on PCR results were disussed by single factor experiments.[Result] The optimized amplification procedure was as follows: pre-denaturing at 94 ℃ for 2 min;denaturing at 94 ℃ for 30 s,annealing at 48 ℃ for 30 s,extending at 72 ℃ for 1 min,40 cycles;extending at 72 ℃ for 7 min.The optimum reaction system was as follows: 30 ng DNA template,2.0 μmol/L upstream primer and downstream primer,0.3 mmol/L dNTPs,2.5 mmol/L Mg2+,the total volume was 25 μl.[Conclusion] The optimization amplification system for SRAP-PCR of ginseng was set up,which provided rapid and simplified test methods with good repeatability for SRAP analysis of the genetic relationship and genetic diversity of ginseng.
Keywords:SRAP  PCR
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