文献类型：期刊文献

英文题名：Isolation of high-quality RNA from Platycladus orientalis and other Cupressaceae plants

作者：Chang, Ermei[1] Zhao, Yuxin[1] Wei, Qianchun[1] Shi, Shengqing[1] Jiang, Zeping[1]

第一作者：常二梅

通信作者：Jiang, ZP[1]

机构：[1]Chinese Acad Forestry, Res Inst Forestry, State Key Lab Tree Genet & Breeding, Beijing, PR, Peoples R China

年份：2016

卷号：23

起止页码：21-27

外文期刊名：ELECTRONIC JOURNAL OF BIOTECHNOLOGY

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000389595000004)】；

基金：This work was supported by the National Natural Science Foundation of China [grant number 31300555]; and the National Non-Profit Research Institutions of Chinese Academy of Forestry [grant numbers RIF2013-12, RIF2014-11, CAFYBB2014QB004].

语种：英文

外文关键词：Cupressaceae plants; Polysaccharides; Polyphenols; Total RNA isolation; Two-step removal of polyphenols and polysaccharides

摘要：Platycladus orientalis has a lifespan of several thousand years in China, making it a good plant in which to study aging at the molecular level, but this requires sufficient quantities of high-quality P. orientalis RNA. However, no appropriate methods have been reported for total RNA isolation from P. orientalis leaves. The TRIzol method did not extract RNA, while cetyltrimethylammonium bromide, sodium dodecyl sulfate-phenol, and plant RNAout kit (Tianz, Inc., China) protocols resulted in low yields of poor quality RNA. Isolating total RNA using the Spectrum (TM) Plant Total RNA Kit (Sigma-Aldrich, St. Louis, MO, USA) resulted in a high-quality product but a low yield. However, the two-step removal of polyphenols and polysaccharides in the improved plant RNAout kit protocol resulted in the isolation of RNA with a 28S: 18S rRNA ratio of band intensities that was similar to 2: 1, the A260/A280 absorbance ratio was 2.03, and the total RNA yield from P. orientalis leaves was high. This protocol was tested on different P. orientalis tissues of different ages and on leaves of five other Cupressaceae plants. The total RNAs were successfully used in complementary DNA synthesis for transcriptome sequencing and would be suitable to use in additional experiments. The results of this study will benefit future studies in Cupressaceae plants. (C) 2016 Pontificia Universidad Catolica de Valparaiso. Production and hosting by Elsevier B.V. All rights reserved.