| 花椰菜RAPD-PCR反应体系的优化 |
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| 引用本文： | 陈阳,周先治,吴宇芬,陈晟,赵依杰.花椰菜RAPD-PCR反应体系的优化[J].江西农业学报,2010,22(11). |
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| 作者姓名： | 陈阳 周先治 吴宇芬 陈晟 赵依杰 |
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| 基金项目： | 福建省科技厅重点项目，福建省自然基金 |
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| 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0.8~1.6μmol/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。
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| 关 键 词： | 花椰菜 随机扩增多态性DNA 正交法 |
Optimization of RAPD-PCR Reaction System in Cauliflower |
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| CHEN Yang,ZHOU Xian-zhi,WU Yu-fen,CHEN Sheng,ZHAO Yi-jie.Optimization of RAPD-PCR Reaction System in Cauliflower[J].Acta Agriculturae Jiangxi,2010,22(11). |
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| Authors: | CHEN Yang ZHOU Xian-zhi WU Yu-fen CHEN Sheng ZHAO Yi-jie |
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| Abstract: | 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. |
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| Keywords: | Cauliflower RAPD Orthogonal experiment Reaction system Optimization |
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