title: Fluorescence labeling of short RNA by oxidation at the 3'-end.
abstract: In RNA nanotechnology, construction of nanoparticles involves conjugation of functionalities, cross-linking of modules, labeling of RNA subunits, and chemical modification of nucleotides. Efficiency and sensitivity are important for the RNA labeling, which also can be used as probes in microarrays, Northern blotting, and gel-shift assays. Here, we describe a method for fluorescence labeling of short RNA at the 3'-end by oxidation. The 3'-terminus of in vitro-transcribed short RNA is oxidized by sodium periodate, and fluorescein-5-thiosemicarbazide is added after removal of excess oxidant. Purified short RNA with fluorescence is then applied for detection of RNA-protein interaction by gel-shift assay.
authors.affiliation: Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 320 Yueyang Road, 200031, Shanghai, China.
authors.forename: Chen
authors.initials: C
authors.lastname: Qiu
authors.forename: Wang-Yi
authors.initials: WY
authors.lastname: Liu
authors.forename: Yong-Zhen
authors.initials: YZ
authors.lastname: Xu
mesh_terms: D019793:Fluorescein / Q000737:chemistry; D005453:Fluorescence; D053758:Nanoparticles / Q000737:chemistry*; D036103:Nanotechnology / Q000379:methods*; D010084:Oxidation-Reduction; D010504:Periodic Acid / Q000737:chemistry; D012313:RNA / Q000737:chemistry* / Q000235:genetics; D012663:Semicarbazides / Q000737:chemistry; D013194:Staining and Labeling
chemical_list: D012663:Semicarbazides; D010504:Periodic Acid; C005151:thiosemicarbazide; D012313:RNA; C009288:metaperiodate; D019793:Fluorescein