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目的:为了得到高效的文心兰RNA提取方法和高质量的RNA,为后续文心兰分子生物学研究奠定基础。方法:选取文心兰“黄金2号”(Oncidium Gower Ramsey‘Gold2’)叶片和根组织为材料,对SDS-LiCl法、改良CTAB-NaAC法和改良CTAB-LiCl法和总RNA提取效果进行了比较研究。结果:改良CTAB-LiCl法得到的RNA样品纯度较高,完整性好,经电泳检测条带清晰无明显降解,28S条带的亮度是18S条带亮度的2倍,从叶片和气生根组织中提取RNA的OD260/OD280比值分别为1.797和1.787,提取率分别为33.07μg/g、29.07μg/g。以此RNA为模板进行RT-PCR反应,能获得特异条带。结论:改良CTAB-LiCl法是一种高效的文心兰RNA提取方法,所得样品RNA适合进一步的分子生物学研究。
OBJECTIVE: To establish an efficient method for the extraction of high-quality RNA from Atractylodes macrocephala and to lay the foundation for the follow-up study of molecular biology. Methods: The leaves and root tissues of Oncidium Gower Ramsey’Gold2 ’were selected as experimental materials to study the effects of SDS-LiCl method, modified CTAB-NaAC method and modified CTAB-LiCl method and total RNA extraction Conducted a comparative study. Results: The RNA samples obtained by the modified CTAB-LiCl method showed higher purity and better integrity. The bands of 28S bands were twice as bright as those of the 18S bands, The OD260 / OD280 ratios of RNA were 1.797 and 1.787 respectively, the extraction rates were 33.07μg / g and 29.07μg / g, respectively. Using this RNA as a template for RT-PCR reaction, specific bands can be obtained. Conclusion: The modified CTAB-LiCl method is an efficient method for the extraction of A. sinensis RNA. The obtained sample RNA is suitable for further molecular biology research.