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以新疆野苹果叶片为试材,采取改良CTAB法提取基因组,参照基本PCR反应体系,采用控制变量的方法,研究了各成分不同的浓度梯度对新疆野苹果SSR-PCR扩增的影响,为新疆野苹果的遗传育种奠定基础。结果表明:新疆野苹果SSR扩增的最佳体系为10×PCR buffer 2.5μL,dNTP 0.2mmol·L~(-1),0.2mmol·L~(-1)引物,0.5UTaq DNA聚合酶,100ng DNA模板。并用此优化体系对154份新疆野苹果进行了稳定性、普遍性验证,扩增效果较好。
Xinjiang wild apple leaves were taken as experimental materials, the improved CTAB method was used to extract the genomes, and the basic PCR reaction system was used. The control variables were used to study the effect of different concentration gradients of each component on the SSR-PCR amplification of Xinjiang apple. Wild apple genetic breeding laid the foundation. The results showed that the optimal system for Xinjiang SSR amplification was 2.5 × 10 × PCR buffer, 0.2 mmol·L -1 dNTP, 0.2 mmol·L -1 primer, 0.5 mmol DNA polymerase, 100 ng DNA template. The stability and universality of 154 wild apple cultivars in Xinjiang were validated by this optimized system, and the amplification effect was better.