黄连药材DNA提取及RAPD反应体系的优化

DNA Extraction from Rhizoma Coptidus and optimization of RAPD reaction system

目的探索黄连药材DNA的提取并对其RAPD反应体系进行优化。方法采用目前较为流行的苯酚法、CTAB法、高盐低pH值法，通过电泳、紫外分光光度仪检测及RAPD扩增效果确定黄连药材基因组DNA提取的最佳方法。结果CTAB法为黄连药材基因组DNA提取的最佳方法，建立了适合黄连药材的RAPD反应体系：25μL体系中内含1×PCR buffer、2mmol／LM g^2＋、100-150μmol／L dNTP、20ng引物、40ng模板、1UTaq酶。结论CTAB法及黄连药材的RAPD反应体系可以用于黄连药材的RAPD分析。

Objective To study the genomic DNA extraction from Rhizoma Coptidus and optimization of RAPD reaction system. Methods Different methods, i.e. phenol method, CTAB method, low pH extraction medium with high salt, were used to genomic DNA extract from Rhizoma Coptidus. The DNA samples obtained by the above methods were tested by agarose gel electrophoresis and ultraviolet spectrometer. Results CTAB Method was considered to be an optimal technique. Based on the genomic DNA extracted by CTAB method, a reaction system suitable for Rhizoma Coptidus was established, that is, 25 μL amplification reactions system containing 1 × PCR buffer, 2 mmol/L Mg^2＋, 100-150 μmol/L dNTP, 20 ng primer, 40 ng template DNA, and 1 U Taq DNA polymerase. Conclusion CTAB Method and RAPD reaction system can be used to RAPD analysis in Rhizoma Coptidus.