从棉铃虫体中提取总RNA的一种有效方法

An Efficient Method of Total RNA Isolation from Helicoverpa armigera

昆虫组织细胞中含有大量的RNA酶 ,在提取其RNA时 ,防止RNA酶的降解 ,是保证所得RNA片段完整的关键。目前提取动物组织细胞总RNA的方法主要采用“异硫氰酸胍 酚 氯仿抽提”一步法操作 ,但从昆虫组织细胞中提取RNA尚无明确的方法。本实验根据昆虫组织细胞中RNA的分子结合其它蛋白等次生物质的特性不同 ,适当的调整该方法 ,从棉铃虫中提取到了完整、无降解、纯度高的RNA 。

The organisms of insects contain large amount of RNAase that can cause extensive degradation by RNA in the samples. It is essential to prevent degradation by RNAase for obtaining intact RNA molecules from these organisms. Now the 'Acid Guanidinium Thiocyanate Phenol Chloroform' (AGPC) has been used in obtaining animals' intact RNA. But there isn't a tangible method in isolating insects' total RNA. In this experiment, we improved this method according to the different molecule characters of RNA and obtained high quality of RNA from Helicoverpa armigera. These RNA are applicable to Northern blotting, cDNA synthesis and other manipulation of molecular biology.