Ouyang Kunxi, Li Juncheng, Huang Hao, Que Qingmin, Li Pei, Chen Xiaoyang
State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangdong Key Laboratory for Innovative Development and Utilization of Forest Plant Germplasm, South China Agricultural University , Guangzhou , Guangdong , P.R. China.
State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangdong Key Laboratory for Innovative Development and Utilization of Forest Plant Germplasm, South China Agricultural University , Guangzhou , Guangdong , P.R. China ; Guangxi Botanical Garden of Medicinal Plants , Nanning , Guangxi , P.R. China.
Biotechnol Biotechnol Equip. 2014 Nov 2;28(6):1008-1013. doi: 10.1080/13102818.2014.981086. Epub 2014 Nov 21.
Plant tissues contain abundant polysaccharides, phenolic compounds and other metabolites, which makes it difficult to isolate high-quality RNA from them. In addition, contains large quantities of other components, particularly RNA-binding alkaloids, which makes the isolation even more challenging. Here, we describe a concise and efficient RNA isolation method that combines the cetyltrimethyl ammonium bromide (CTAB) and Plant RNA Kit (Omega) protocols. Gel electrophoresis showed that RNA extracted from all tissues, using this protocol, was of good integrity and without DNA contamination. Furthermore, the isolated RNA was of high purity, with an / ratio of 2.1 and an / ratio of >2.0. The isolated RNA was also suitable for downstream applications, such as reverse transcription-polymerase chain reaction (RT-PCR) and quantitative RT-PCR (RT-qPCR). The RNA isolation method was also efficient for recalcitrant plant tissues.
植物组织含有丰富的多糖、酚类化合物和其他代谢物，这使得从它们中分离高质量RNA变得困难。此外， 含有大量其他成分，特别是RNA结合生物碱，这使得分离更具挑战性。在这里，我们描述了一种简洁高效的RNA分离方法，该方法结合了十六烷基三甲基溴化铵（CTAB）和植物RNA提取试剂盒（Omega）的方案。凝胶电泳显示，使用该方案从所有组织中提取的RNA完整性良好，且无DNA污染。此外，分离出的RNA纯度高，A260/A280比值为2.1，A260/A230比值>2.0。分离出的RNA也适用于下游应用，如逆转录聚合酶链反应（RT-PCR）和定量RT-PCR（RT-qPCR）。该RNA分离方法对难处理的植物组织也有效。
