| 一种提取高质量油菜种子和种皮RNA的方法 |
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| 引用本文： | 严明理,刘忠松,官春云,陈社员,刘显军.一种提取高质量油菜种子和种皮RNA的方法[J].生物技术通报,2007(6):97-100. |
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| 作者姓名： | 严明理 刘忠松 官春云 陈社员 刘显军 |
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| 作者单位： | 1. 湖南农业大学农学院,长沙,410128;湖南科技大学生命科学院,湘潭,411201
2. 湖南农业大学农学院,长沙,410128 |
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| 基金项目： | 国家自然科学基金(30471098)，教育部重点科技项目(204101)，湖南省教育厅重点项目(03A017) |
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| 摘 要： | 提取高质量的RNA是从基因表达水平上研究油菜种子和种皮发育的必要条件。现有方法因为油菜种子脂肪、多酚和多糖,难以快速获得完整、高纯度的油菜种子总RNA。本试验针对油菜种子和种皮特点,利用苯酚-氯仿抽提后用无水乙醇沉淀RNA,建立了在油菜种子和种皮中快速提取高质量总RNA的提取方法,电泳分析表明28S rRNA亮度约为18S rRNA的2倍;紫外分光光度计检测A260/A280介于1.8～2.0之间。用该法分离的RNA,已成功用于RT-PCR、Northern blot分析和基因全长的克隆等分子生物学研究。
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| 关 键 词： | RNA提取 油菜 种子 油菜种皮 |
| 收稿时间： | 2007-05-21 |
| 修稿时间： | 2007年5月21日 |
A Method for High-quality RNA Isolation from the Seeds and Their Testa of Rapeseed |
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| Yan Mingli Liu Zhongsong Guan Chunyun Chen Sheyuan Liu Xianjun.A Method for High-quality RNA Isolation from the Seeds and Their Testa of Rapeseed[J].Biotechnology Bulletin,2007(6):97-100. |
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| Authors: | Yan Mingli Liu Zhongsong Guan Chunyun Chen Sheyuan Liu Xianjun |
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| Abstract: | Isolation of high-quality RNA is a prerequisite to gene expression analysis of the seeds and their testa in rapeseed.It is difficult to isolate high-quality total RNA from rapeseed rapidly with previous extraction methods because of much polyphenols,Fats and the polysaccharides in rapaseed seed.An efficient method of extracting total RNA has been developed basing on the seed characteristic,RNA was deposited from ethanol after extracting in phenol and chloroform.Electrophoresis showed the ratio of intensity of the 28S band to that of 18S rRNA band was 2:1.The absorbance ratios(A260/A280)of purified RNA were 1.8 to 2.0.The RNA extracted from the seeds and their testa of rapeseed with this method could be used in molecular biological researches such as RT-PCR,Northern blotting analysis and full-length gene cloning. |
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| Keywords: | RNA isolation Rapeseed Seed Seed coat |
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