直立白薇基因组DNA提取条件的优化

Optimization of Genome Extraction FromCynanchum atratum Bge.

以新鲜直立白薇叶片为试材,采用DNA试剂盒提取方法,研究了DNA提取过程中琼脂糖凝胶浓度、样品加样量、洗脱液用量及乙醇浓度4个因素对直立白薇基因组DNA提取效果的影响,以期获得最适宜直立白薇基因组DNA的提取方法。结果表明:琼脂糖凝胶浓度为0.8%,点样量为13μL,同时以80%乙醇沉淀,洗脱缓冲液用量为100μL时电泳条带最为清晰明亮、不拖尾,为直立白薇基因组DNA最优提取方法。综上,基于试剂盒方法进行提取优化,获得高纯度直立白薇DNA,为直立白薇分子生物学及遗传学的研究奠定基础,为保证正品直立白薇药材资源开发和利用提供参考依据。

The fresh leaves of Cynanchum atratum were used as experimental materials,and the DNA kit extraction method was used to study the effects of agarose gel concentration,sample volume,eluent volume and ethanol concentration on the extraction of genomic DNA from Cynanchum atratum,in order to obtain the most suitable extraction method of genomic DNA from Cynanchi atratum.The results showed that when the concentration of agarose gel was 0.8%,the amount of sample was 13μL,80%ethanol was used for precipitation,and the amount of elution buffer was 100μL,the electrophoresis band was the most clear and bright without tailing,which was the optimal method for genomic DNA extraction of Cynanchum atratum.In summary based on the kit method,the high purity of Cynanchum atratum DNA was obtained,which laid the foundation for the study of molecular biology and genetics of Cynanchum atratum,and provided reference for the development and utilization of authentic Cynanchi atratum medicinal resources.