摘要
以岩陀植物嫩叶为材料,用不同方法对岩陀叶片DNA进行提取,比较提取的质量差异,寻找适合岩陀叶片总DNA的提取方法,以满足多基原民族药岩陀DNA条形码识别系统构建的研究需要。采用十六烷基三甲基溴化铵(CTAB)法、改良的CTAB法、十二烷基硫酸钠(SDS)法及碱裂解法提取岩陀叶片总DNA,利用琼脂糖凝胶电泳、紫外分光光度法及PCR扩增效果检查DNA提取的质量。结果显示四种方法均能从岩陀植物叶片中提取到基因组DNA,改良CTAB法提取的DNA纯度和完整性明显好于其他方法,且采用此法进行PCR扩增获得的目标条带明亮单一,扩增效率达到100%。改良的CTAB法是岩陀叶片总DNA的提取中最为合适的提取方法,能够满足后续DNA条形码的研究需要。
Fresh leaves of Rodgersia pinnata were used as experimental materials, the differences of the DNA extraction effects among different methods were compared to find proper DNA extraction methods for meeting the study demands on constructing DNA barcode recognition system of R. pinnata with different origins. CTAB method, improved CTAB method, SDS method and alkaline lysis method were used to extract total DNA, the quality of DNA extraction were evaluated by agarose gel electrophoresis, UV spectrophotometry and PCR amplification effect. The results showed that all four methods could extract genomic DNA from the leaves of R. pinnata, DNA purity and integrity in improved CTAB method is significantly higher than that in other methods, in which the PCR amplification efficiency reaches 100%, and the amplified bands are bright and clear. The improved CTAB method can be used as a suitable method for extracting total DNA from the leaves of R. pinnata and could meet the study demand of DNA barcode.
作者
方强强
王燕
FANG Qiang-qiang;WANG Yan(College of Pharmacology,Dali University,Dali,Yunnan 671000,China)
出处
《井冈山大学学报（自然科学版）》
2019年第2期19-24,共6页
Journal of Jinggangshan University (Natural Science)
基金
国家自然科学基金项目(81560695)
大理大学博士启动基金项目(KYBS201512)
大理大学第八批中青年学术带头人培养基金项目
