青藏高原珍稀藏药水母雪莲药材DNA微量提取方法优化

Optimization of DNA Micro-extraction Method of Saussurea medusa Maxim, A Rare Tibetan Medicinal Plant of Qinghai-Tibet Plateau

本研究采用改良CTAB法、改良SDS法、高盐低p H法提取自然风干的水母雪莲植株的基因组DNA,并对提取方法进行优化筛选出适合于水母雪莲药材DNA的提取方法。研究表明改良CTAB(CTAB浓度为2%,PVP浓度为1%,β-巯基乙醇浓度为2%)法所提取的基因组DNA得率较高,且使用氯仿/异戊醇抽提两次时提取出的DNA质量较好,能够满足ISSR分析要求。在此基础上对实验材料进行预处理,优化获得了完全适用于水母雪莲风干药材DNA提取的一整套提取流程。该方法能够有效去除次生代谢产物对DNA提取的影响,且对原材料消耗较少,可以应用于后续水母雪莲种质资源鉴定和遗传多样性分析。

In this study, Genomic DNA of dried Saussurea medusa was extracted through CTAB-improved method,SDS-improved method and high salt low p H basic method, then optimized and screened a suitable method for DNA extraction of S. medusa. Results showed that the yield of genomic DNA extracted by CTAB-improved method(2%CTAB concentration, 1% PVP concentration, 2% β-mercaptoethanol concentration) was higher than others. And two times of extraction with chloroform/isoamyl alcohol had got the better quality of DNA that met the requirements of ISSR analysis. On the basis of this, preprocessed the experimental materials and got a complete set of DNA extraction process of S. medusa dried herbs. This improved method can effectively remove the effect of secondary metabolites and consume less plant materials and it also can be applied to the germplasm identification and genetic diversity analysis of S. medusa.