摘要
用改进后的SDS法从夏枯草植物的新鲜叶片中提取基因组DNA,通过正交设计和单因素结合的方法对RAPD-PCR反应条件进行优化,确定了适合夏枯草DNA的最佳扩增体系:20μL的PCR反应体系中,模板DNA浓度1.0-2.0 mg/L,引物浓度1.00-1.50 mmol/L,Mg2+浓度2.0-2.5 mmol/L,dNTP浓度0.20-0.25 mmol/L,Taq酶用量0.5 U,10×BufferMg2+free 2μL,BSA浓度0.25-0.50μg/μL。
An improved SDS method of DNA extraction is applied to obtaining genomie DNA from the fresh leaves of Prunella vulgaris L.. In this paper, we used orthogonal experiment combining with single factor experiment to optimize the RAPD reaction conditions. The result showed that the optimal conditions in 20 μL PCR system were DNA 1.0 ～ 2.0 mg/L, random primer 1.00 ～ 1.50 μmol/L, Mg^2＋ 2.0 ～ 2.5 mmol/L, dNTP 0. 20 ～ 0. 25 mmol/L, Taq polymerase 0. 5 U, 10 × Buffer Mg^2＋ free 2 μL, BSA 0. 25 ～ 0. 50 μg/μL.
出处
《天然产物研究与开发》
CAS
CSCD
2007年第1期123-126,134,共5页
Natural Product Research and Development
