基于单因素试验的大豆CDDP-PCR反应体系优化  

Optimization of CDDP-PCR System by Single factor Experiment Design in Soybean

作　　者：李强 高聚林[2] 苏二虎 廉博[3] 牛敏 邵志壮 谢田 

机构地区：[1]内蒙古自治区农牧业科学院,内蒙古呼和浩特010031 [2]内蒙古农业大学农学院,内蒙古呼和浩特010019 [3]呼伦贝尔市农业技术推广服务中心,内蒙古呼伦贝尔021008 [4]呼和浩特市种子管理站,内蒙古呼和浩特010020

出　　处：《安徽农业科学》2015年第3期32-34,43,共4页Journal of Anhui Agricultural Sciences

基　　金：内蒙古自治区科技创新引导奖励资金项目(20111707);内蒙古农牧业科学院青年创新基金项目(2014QNJJN01)

摘　　要：[目的]优化大豆CDDP-PCR反应体系。[方法]采用单因素试验方法对影响大豆CDDP-PCR反应的Mg2+浓度、Taq DNA聚合酶用量、引物浓度、d NTPs浓度和模板DNA用量等因素进行优化。[结果]优化后的大豆CDDP-PCR体系为:Mg2+浓度2.0 mmol/L、Taq聚合酶用量1.5 U、引物浓度0.375μmol/L、d NTPs浓度0.3 mmol/L、DNA模板用量40 ng。运用24个大豆品种验证了该反应体系稳定、可靠。[结论]该反应体系的建立为大豆种质遗传多样性分析、遗传连锁图谱构建及分子标记辅助育种奠定了基础。[Objective]The aim was to optimize CDDP-PCR system by single factor experiment design in soybean. [Method]Single factor experiment design were applied to optimize CDDP-PCR amplification system of soybean in five factors such as Mg2 ＋,Taq DNA polymerase,primer,d NTPs and template DNA. [Result]The suitable reaction system was obtained,that is 20 μl containing 2. 0 mmol / L Mg2 ＋,1. 5 U Taq polymerase,0. 375 μmol / L primer,0. 3 mmol / L d NTPs,40 ng DNA template. The optimized CDDP-PCR system was tested on 24 soybeans,and the result was stable and reliable. [Conclusion]The system provides the basis for evaluation of genetic diversity,construction of genetic linkage map and molecular marker assisted breeding for the soybean.

关 键 词：大豆 CDDP 单因素试验 体系优化 

分 类 号：S565.1[农业科学—作物学]