草坪草ctDNA提纯方法探讨

摘要

以4种草坪草高羊茅Festuca elata、黑麦草Lotium perenne、狗牙根Cynodon dactylon、中华结缕草Zoysia sinica为材料,用改进的差速离心法分离叶绿体, CTAB法提取叶绿体DNA(ctDNA),琼脂糖电泳检测ctDNA含量.结果显示,改进CTAB法提取ctDNA的纯度高、谱带清晰、完整叶绿体比例高;且CTAB法与常用高等植物ctDNA的分离提纯方法相比,省去传统的密度梯度离心法耗材、耗时步骤,具有简便、快速、得率较高等优点.实验表明,改进CTAB法为禾本科植物ct DNA的提取提供参考,适于生物实验教学和科研活动.%Four turfgrass including Festuca elata, Lotium perenne, Cynodon dactylon and Zoysia sinica were selected as the materials, the chloroplast was isolated by improved differential centrifugation, and the ctDNA was extracted with the CTAB and tested by Agarose gel electrophoresis. The results showed that the purity of ctDNA was high, the bands were clear, the ratio of intact chloroplast was high with the improved CTAB method.Compared with traditional method of ctDNA isolation and purification, CTAB method eliminated the steps of consuming material and time in traditional density gradient centrifugation; it was simple, rapid and high yield. So the improved CTAB method provided a reference for the extraction of Gramineae plants ctDNA, which was applicable for biological experimental teaching and research activities.