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本文用水稻单粒干种子作材料 ,进行了 DNA提取方法和 RAPD程序优化研究。结果表明 :CTAB法微量提取水稻干种子 DNA可获得理想的扩增结果 ,最佳的反应体系和条件是 :随机引物浓度 1 0 0～ 40 0 μg/L、Mg2 + 浓度 2～ 4mmol/L、d NTPs浓度 1 0 0～ 40 0 μmol/L、Taq酶 1 U。变性 94℃ 3 0 s,退火 3 8℃ 3 0 s,延伸 72℃ 1 min,40个循环。为了节约时间和成本 ,可不进行予变性处理 ,反应体积可降至 1 2 .5μl,扩增效果一样。
In this paper, a single dry seed of rice as materials, DNA extraction methods and RAPD program optimization. The results showed that the optimal reaction conditions were as follows: the concentration of random primers was 100 ~ 40 0 μg / L, the concentration of Mg2 + was 2 ~ 4 mmol / L, d NTPs concentration of 100 ~ 40 0 μmol / L, Taq enzyme 1U. Denaturation at 94 ℃ for 30 s, annealing at 38 ℃ for 30 s and extension at 72 ℃ for 1 min for 40 cycles. In order to save time and cost, without pre-denaturation, the reaction volume can be reduced to 12.5μl with the same amplification effect.