一种用于黄曲霉高质量总RNA提取方法与应用  

A Method for Total RNA Extraction from Fungus Aspergillus flavus and Its Application

作　　者：林剑青[1] 王成成[1] 肖春华[1] 陈国华[1] 贺竹梅[1] 

机构地区：[1]中山大学生命科学学院水产品安全教育部重点实验室,广东广州510275

出　　处：《中山大学学报（自然科学版）》2012年第6期107-111,共5页Acta Scientiarum Naturalium Universitatis Sunyatseni

基　　金：国家自然科学基金资助项目(31170044)

摘　　要：不断发展的研究技术对黄曲霉RNA的提取质量提出更高的要求,而真菌的细胞壁结构及其内源性RNase活性常是达到这一目标的主要障碍。以黄曲霉菌丝为材料,探讨了采用TES热酚法获得高质量总RNA的方法,并通过RT-PCR(反转录聚合酶链式反应)和RNA-Sequencing(转录组测序)对获得的RNA进行了质量验证。结果显示,TES热酚法不但所用试剂价格低廉,该法所提取的RNA质量较高,其浓度、纯度和完整性均可满足RT-PCR、转录组测序的文库构建和高通量测序等对RNA要求较高的分子生物学实验。The rapidly developing molecular techniques require the ability to extract high quality RNA from Aspergillus flavus. The cell wall structure and endogenous RNase activity in A. flavus are often the main obstacles for obtaining RNA with high quality. In this paper, TES hot phenol method was applied to obtain total RNA from A. flavus, and the total RNA was further used in RT-PCR （ Reverse translation-polymerase chain reaction） and RNA-sequencing. The results showed that the TES hot phenol method can be used to get a good quality of total RNA from this filamentous fungus as templates for following studies, such as RT-PCR and RNA sequencing.

关 键 词：黄曲霉 RNA提取 TES热酚法 RT—PCR 转录组测序 

分 类 号：Q78[生物学—分子生物学]