摘要
目的优选胡黄连稳定的ISSR-RCR反应体系。方法采用4因素(dNTRs、Mg2+、Taq酶、引物)4水平的正交实验和单因子梯度优化相结合的方法。结果适于胡黄连的25μl ISSR-PCR反应体系中各因素最佳浓度分别为10×buffer2.5μl,dNTPs 150μmol/L,Mg2+1.5mmol/L,Taq酶1U,引物0.5μmol/L,DNA20 ng或10×buffer 2.5μl,dNTPs 150μmol/L,Mg2+2 mmol/L,Taq酶0.5 U,引物0.4μmol/L,DNA20 ng。结论对于胡黄连ISSR-RCR试验,一次正交实验完全可以建立起满足要求的PCR体系。
Objective To optimize the stable ISSR reaction system for Neopicrorhiza scrophulariiflora. Methods Concentration of dNTPs,Mg^2＋ , Taq DNA polymerase and primers were studied by orthogonal experiment combined with single factor experiment. Results The optimal ISSR - PCR reaction system（25μl） was as follows ： 10 x buffer 2.5 μl, dNTPs150 μmol/L, Mg2 ＋ 1.5mmol/ L, Taq DNA polymerase 1U,primers 0.5 μmol/L, DNA 20 ng or 10× buffer 2.5 μl, dNTPs 150 μmoL/L, Mg^2＋ 2mmol/L,Taq DNA polymerase 0.5U ,primers 0.4μmol/L, DNA 20 ng. Conclusion For Neopicrorhiza scrophulariiflora, one orthogonal experiment can establish satisfactory PCR system.
出处
《时珍国医国药》
CAS
CSCD
北大核心
2010年第2期323-325,共3页
Lishizhen Medicine and Materia Medica Research
基金
国家科技基础性工作专项重点项目子课题(No.SB2007FY0200)
云南中医学院科学研究基金项目(No.200811)
关键词
胡黄连
正交实验
单因子
ISSR—PCR
Neopicrorhiza scrophulariiflora
Orthogonal experiment
Single factor experiment
ISSR- PCR
