title: An Efficient Method for the Isolation of Highly Purified RNA from Seeds for Use in Quantitative Transcriptome Analysis.
abstract: Plant seeds accumulate large amounts of storage reserves comprising biodegradable organic matter. Humans rely on seed storage reserves for food and as industrial materials. Gene expression profiles are powerful tools for investigating metabolic regulation in plant cells. Therefore, detailed, accurate gene expression profiles during seed development are required for crop breeding. Acquiring highly purified RNA is essential for producing these profiles. Efficient methods are needed to isolate highly purified RNA from seeds. Here, we describe a method for isolating RNA from seeds containing large amounts of oils, proteins, and polyphenols, which have inhibitory effects on high-purity RNA isolation. Our method enables highly purified RNA to be obtained from seeds without the use of phenol, chloroform, or additional processes for RNA purification. This method is applicable to Arabidopsis, rapeseed, and soybean seeds. Our method will be useful for monitoring the expression patterns of low level transcripts in developing and mature seeds.
authors.affiliation: Laboratory of Biological Diversity, Department of Evolutionary Biology and Biodiversity, National Institute for Basic Biology.
authors.forename: Masatake
authors.initials: M
authors.lastname: Kanai
authors.affiliation: Laboratory of Biological Diversity, Department of Evolutionary Biology and Biodiversity, National Institute for Basic Biology; Department of Basic Biology, SOKENDAI (The Graduate University for Advanced Studies); mano@nibb.ac.jp.
authors.forename: Shoji
authors.initials: S
authors.lastname: Mano
authors.affiliation: Department of Cell Biology, National Institute for Basic Biology.
authors.forename: Mikio
authors.initials: M
authors.lastname: Nishimura
mesh_terms: D017360:Arabidopsis / Q000235:genetics; D029741:Brassica rapa / Q000235:genetics; D020869:Gene Expression Profiling; D018749:RNA, Plant / Q000302:isolation & purification*; D012639:Seeds / Q000235:genetics*; D013025:Glycine max / Q000235:genetics; D059467:Transcriptome
chemical_list: D018749:RNA, Plant