文心兰RNA不同提取方法比较

Comparison of Different RNA Extraction Methods of Oncidium Orchid

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： In order to obtain efficient extraction method and high quality of RNA from Oncidium orchid for subsequent molecular research. Method： Comparison of SDS - LiC1 method, modified CTAB - NaAC and modified CTAB - LiC1 method for total RNA extraction from roots and leaves of Oncidium Cower Ramsey ＇ Cold2＇ was conducted. Result： The RNA samples obtained by the modified CTAB - LiC1 method were of higher purity, integrity and no degradation. As detected with OD value and gel electrophoresis, bands were clearly visi- ble ,the band of 28S was 2 times bright that of 18S rRNA,OD260/OD280 ratios of RNA from leaves and gas roots were 1. 797 and 1. 787 ,and extraction ratios were 33.07 and 29.07, respectively. With the RNAs as templates for RT - PCR reaction, specific bands were produced. Conclusion：The modified CTAB - LiC1 method is an efficient extraction method of RNA from Oncidium orchid and the RNA samples obtained could be used for subsequent molecular biological operations.