Kolosova Natalia, Miller Barbara, Ralph Steven, Ellis Brian E, Douglas Carl, Ritland Kermit, Bohlmann Jörg
University of British Columbia, Vancouver, BC, Canada.
Biotechniques. 2004 May;36(5):821-4. doi: 10.2144/04365ST06.
An improved protocol was developed for efficient and reliable extraction of high-quality total RNA and mRNA from various tissues of spruce (Picea spp.) and poplar (Populus spp.) trees, as well as other plant species. This method was specifically optimized for tissues with high content of polysaccharides, oleoresin terpenoids, and phenolic secondary metabolites, which often co-precipitate with RNA and inhibit subsequent reverse transcription. The improved protocol yielded up to 600 micrograms of total RNA per gram of tissue suitable for standard expressed sequence tags (ESTs), full-length cDNA library construction, and for microarray applications.
开发了一种改进的方案,用于从云杉(Picea spp.)和杨树(Populus spp.)树以及其他植物物种的各种组织中高效、可靠地提取高质量的总RNA和mRNA。该方法针对多糖、油树脂萜类化合物和酚类次生代谢物含量高的组织进行了专门优化,这些物质常与RNA共沉淀并抑制后续的逆转录。改进后的方案每克组织可产生高达600微克的总RNA,适用于标准表达序列标签(EST)、全长cDNA文库构建和微阵列应用。