甘蓝型油菜籽粒RNA提取方法的探讨

Research on the Methods of Isolation of RNA from Oilseeds(Brassica napus. L)

用CTAB法、SDS法、上海华瞬植物叶RNA抽提试剂盒分别提取甘蓝型油菜花后40 d的籽粒RNA。用电泳法和A260/A280与A260/A280的比值比较提取RNA的质量。结果发现:用SDS法、上海华瞬植物叶RNA抽提试剂盒都没有成功提取RNA,只有CTAB法成功地提取出了完整的甘蓝型油菜籽粒RNA。用CTAB法提取的总RNA的A260/A230与A260/A280的值分别为2.06和1.94,说明采用该方法提取的RNA污染小,纯度较高,获得的RNA产量也较高。通过逆转录实验和基因片段的克隆也证明获得的RNA质量比较高,可以直接满足一般的分子实验。

Based on the methods of CTAB, SDS and Plant （Leaves） Total RNA Isolation Kit, total RNA was isolated from seeds of 40 days of flower from Braccica napus. L. The total RNA was compared by gel electrophoresis and the ratio of A260/A280 and A260/A230. RNA only was separated successfully from the seeds by the methods of CTAB. The sharpness of 28S bands was twice than 18S of bands and the ratio of A260/A280 and A260/A230 of RNA was 1.94 and 2. 06, respectively. It indicated the quality of RNA was high, reverse transcription proved the quality of RNA was fine enough for ecumenical experimentation.