摘要
目的优化味连简单序列重复间扩增多太性(Coptis chinensisFranch.ISSR)反应体系。方法利用正交实验设计,从Mg2+,dNTP,引物,BSA这4种因素3个水平进行优化。结果确立了适合黄连ISSR分析的反应体系,即在25μl反应体系中,内含1×PCR buffer,1.5 mmol/LMg2+,200μmol/L dNTP,0.4μmol/L引物,40 ng模板,1UTaqDNA聚合酶,BSA 1μg/μl。结论利用此优化反应体系能得到清晰、稳定的图谱。
Objective To optimize ISSR amplification system on Coptis chinensis Franch. Methods Orthogonal design was used to investigate four factors（Mg^2＋, dNTP, primer, BSA） at three levels respectively. Results A reaction system suitable for Coptischinensis Franch. was established , that was, 25 μl amplification reactions system containing 1 x PCR buffer, 1.5 mmol/L Mg^2 ＋ , 200 μmol/L dNTP,0.4 μmol/L primer,40 ng template DNA, 1U Taq DNA polymerase, BSA 1 μg/μl. Conclusion A reliable graph can be obtained through this optimal reaction system .
出处
《时珍国医国药》
CAS
CSCD
北大核心
2008年第1期11-12,共2页
Lishizhen Medicine and Materia Medica Research
基金
重庆市科技计划资助项目(No.8847)
国家科技攻关计划(No.2004BA721A32)
国家中医药管理局攻关项目资助(No.2004ZX06-1)
