CAJ | 학술논문

[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、Mg2+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2.0 μmol/L,dNTP浓度 0.3 mmol/L,Mg2+2.5 mmol/L,总体积25 μl.[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.
[목적]연구인삼기인조SRAP-PCR 적확증조건,건립기우화확증체계.[방법]채용단인자실험방법탐토모판DNA、인물농도、dNTPs농도、Mg2+농도등인소대PCR 결과적영향.[결과]우화후적확증정서위:94 ℃ 예변성2 min;94 ℃ 변성30 s,48 ℃ 복성30 s,72 ℃ 연신1 min,공40차순배;72 ℃연신7 min.최가반응체계위:DNA모판30 ng,상하유인물농도2.0 μmol/L,dNTP농도 0.3 mmol/L,Mg2+2.5 mmol/L,총체적25 μl.[결론]건립료만족인삼SRAP-PCR 적우화확증체계,위인삼친연관계화유전다양성SRAP분석제공쾌속、간편、중복성호적실험방법.
[Objective] The research aimed study the amplification condition of SRAP-PCR of ginseng genome and set up the optimized amplification system/ [Method] The effects of template DNA, primer, dNTP mixture and Mg2+ on PCR results were discussed. [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 and 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 μM upstream primer and downstream primer, 0.3 mM dNTP mixture, 2.5 Mm 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.