RNA molecules mature through various posttranscriptional modifications and editing. To characterize base modifications and terminal chemical structures of fully processed native RNAs, it is necessary to isolate individual RNA species from a limited quantity and complex mixture of cellular RNAs. However, there have been no general and convenient strategies for isolation of individual RNAs. We describe a simple and practical method for effective isolation of multiple individual RNA species named "Chaplet Column Chromatography (CCC)." We successfully isolated all species of mammalian mitochondrial tRNAs with this method.