摘要
采用正交法对影响花椰菜RAPD-PCR反应的5个实验因素进行了优化,结果表明,花椰菜RAPD反应的优化体系为:dNTPs 0.12 mmol/L.引物0.8～1.6 μmol/L,模板DNA为20～80 ng,Taq酶为0.75 U,退火温度34～36℃.试验证明,Taq酶用量对RAPD-PCR反应体系的影响最大,退火温度、引物浓度、dNTPs用量对RAPD-PCR反应结果影响均小于Taq用量,但相差不大.模板浓度的影响最小.
Five essential factors that might affect the result of RAPD-PCR reaction of cauliflower were optimized by applying orthogonal experiment.The results showed that the optimal conditions for RAPD-PCR reaction system of cauliflower were dNTPs 0.12 mmol/L,random primer 0.8~1.6 μmol/L,DNA template 20~80 ng,Taq polymerase 0.75 U,annealing temperature 34~36 ℃.Taq enzyme was the most important factor influential to the RAPD-PCR reaction system among the above five essential factors,followed by annealing temperature,primer concentration and dNTPs dosage,and the concentration of template had the lest influence on the RAPD-PCR reaction system.
出处
《江西农业学报》
CAS
2010年第11期5-7,共3页
Acta Agriculturae Jiangxi
基金
福建省科技厅重点项目"台湾十字花科
葫芦科蔬菜良种引进筛选及栽培技术研究"(2007I0037)
福建省自然基金"镧对西瓜光合系统能量传递和强光防御机制调节作用的研究"(2009J01069)
