【摘要】 以短柄(Quercus glanduliferavar.brevipetiolata)的DNA为材料,对影响PCR扩增效果的一些因素诸如模板DNA用量、Taq酶的用量、镁离子浓度、dNTP的浓度、引物用量和牛血清白蛋白浓度等指标进行筛选和优化。结果显示适合短柄ISSR-PCR分析最适宜的PCR反应条件为:10μL PCR反应体积中,1×Taq酶配套缓冲液(10 mmol.L-1Tris-HCl,pH9.0,50 mmol.L-1KCl,0.1%Triton X-100),0.75UTaq DNA聚合酶,2 mmol.L-1MgCl2,0.2 mmol.L-14×dNTP,12 pmol引物,10 ng模板DNA,2 mg.mL-1牛血清白蛋白。短柄ISSR扩增较适宜的退火温度为54.4℃。更多还原

【Abstract】 Factors which affect the ISSR-PCR amplification such as template DNA usage,Taq DNA polymerase,Mg²⁺ concentration,dNTP concentration,primer usage and bovine serum albumin concentration,were optimized and selected by using the materials of Quercus glandulifera var.brevipetiolata DNA.The results showed that the conditions suitable for ISSR-PCR of Quercus glandulifera var.brevipetiolata were as follows:1×Taq buffer(10 mmol·L^-1 Tris-HCl,pH9.0,50 mmol·L^-1 KCl,0.1% Triton X-100),0.75U Taq DNA polymerase,2.0 mmol·L^-1 MgCl₂,0.2 mol·L^-1 4×dNTP,12 pmol primers,10ng template DNA、2 mg·mL^-1 bovine serum albumin. The suitable annealing temperature in the ISSR-PCR reaction system was 54.4℃.更多还原