【摘要】 采用CTAB法提取大白菜基因组DNA,利用单因素试验对InDel-PCR反应体系中的dNTPs浓度、Mg2+浓度、Taq DNA聚合酶用量、模板DNA用量、引物浓度、退火温度、三温循环时间等7个因素进行筛选和优化。结果表明,适用于大白菜InDel-PCR分析的最佳扩增条件为(20μL):dNTPs 0.1mmol/L、Mg2+1.5mmol/L、Taq DNA聚合酶0.4U、模板DNA 105ng、引物浓度0.4μmol/L、退火温度50℃、三温循环时间30s-30s-45s。该体系可用于大白菜遗传多样性分析及遗传图谱构建。更多还原

【Abstract】 The genomic DNA of Chinese cabbage(Brassica rapa L.ssp.pekinensis) was isolated using the CTAB method.Seven factors(dNTP,Mg2+,Taq DNA polymerase,template DNA,primer,annealing temperature,and three temperature zones for PCR reaction) affecting InDel-PCR reaction system were optimized using a single factor experiment.The electrophoresis showed clear bands,which was applied to analyze Brassica rapa InDel-PCR amplification conditions.The optimal amplification conditions(20 μL) were:dNTP 0.1 mmol/L,Mg2+ 1.5mmol/L,Taq DNA polymerase 0.4 U,template DNA 105 ng,primer 0.4 μmol/L,annealing temperature 50 ℃,and three temperature zones 30 s-30 s-45 s.It can be used in genetic diversity analysis and genetic map construction of Chinese cabbage.更多还原