【摘要】 以"嘎啦"苹果组培苗叶片诱导的愈伤组织作为DNA模板提取材料,采用正交设计,摸索了SSR反应体系中Mg²⁺、Taq DNA聚合酶、引物、模板DNA、dNTP的最适浓度,可为苹果愈伤组织的遗传变异研究奠定基础。结果表明:各因素不同水平变化对反应体系的影响为Taq DNA聚合酶>dNTP用量>Mg²⁺用量>引物用量=模板DNA浓度。确立了苹果愈伤组织SSR-PCR最佳反应体系总体积25μL,其中4μL 25mmol·L^-1 Mg²⁺,0.25μL 5U·μL^-1 Taq DNA聚合酶,3μL 0.01mmol·L^-1 SSR引物,1μL 30ng·μL^-1模板DNA,4μL 2.5mmol·L^-1dNTP,2.5μL 10×PCR Buffer,10.25μL超纯水。更多还原

【Abstract】 In order to establish the basis of genetic variation research of apple callus,the ‘Gala’induction of callus was DNA template extraction materials.the Mg²⁺,Taq DNA polymerase,primer dosage,template DNA concentration and dNTP which were suitable for concentration or dosage were explored.The results showed that the effect of different levels on reaction system was Taq DNA polymerase>dNTP dosage>Mg²⁺dosage>primer dosage=template DNA concentration.The optimal reaction system of apple callus was established,the optimal reaction system of apple callus was that25μL for total volume,which were 4μL 25 mmol·L^-1 Mg²⁺,0.25μL 5 U ·μL^-1 Taq DNA polymerase,3μL 0.01 mmol·L^-1 SSR,1μL 30 ng·μL^-1 template DNA,4μL 2.5 mmol·L^-1 dNTP,2.5μL 10×PCR Buffer,10.25μL ultra-pure water.更多还原