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本文以黄早四、汶黄、Mo17等我国主要玉米自交系为材料,使用国产PCR仪,对玉米RAPD分析中PCR过程的DNA模板浓度,扩增反应的循环数进行了研究,并针对国产PCR扩增仪(PCR—90AD)的特点,确定了DNA变性,引物和模板DNA结合,引物沿模板延伸3个步骤的时间。实验结果表明玉米PCR过程使用10～15ng的模板DNA,在94℃模板变性时间1分30秒,36℃引物退火2分钟,72℃引物延伸2分30秒,扩增循环35～40周期条件下扩增效果较好,使用这一RAPD程序研究不同自交系间遗传差异,结果与系谱法基本一致,而同一自交系的10个单株之间,RAPD带型是一致的。本实验建立了适合玉米RAPD分析的PCR程序及条件,为RAPD分析法应用于玉米的遗传研究打下了良好的基础。
In this paper, we studied the concentration of DNA template and the number of cycles of amplification reaction in the RAPD analysis of maize using the domestic main inbred lines of Maize as early as Huangzao 4, Wenhuang and Mo17, The PCR Amplifier (PCR-90AD) is characterized by DNA denaturation, primer-template DNA binding, and primer extension along the template for three steps. The experimental results showed that 10-15ng template DNA was used in the PCR of maize, with the time of denaturation at 94 ℃ for 1 minute and 30 seconds, the primer annealing at 36 ℃ for 2 minutes, the primer extension at 72 ℃ for 2 minutes and 30 seconds and the amplification cycle at 35-40 cycles The amplification effect was good. Using this RAPD program to study the genetic differences among different inbred lines, the results were basically the same as the pedigree method. However, the RAPD bands were consistent among 10 plants of the same inbred line. This experiment established a PCR procedure and conditions suitable for RAPD analysis of maize, which laid a good foundation for RAPD analysis applied to maize genetic research.