title: Four-leaf clover qRT-PCR: A convenient method for selective quantification of mature tRNA.
abstract: Transfer RNAs (tRNAs) play a central role in translation and also recently appear to have a variety of other functions in biological processes beyond translation. Here we report the development of Four-Leaf clover qRT-PCR (FL-PCR), a convenient PCR-based method, which can specifically quantify individual mature tRNA species. In FL-PCR, T4 RNA ligase 2 specifically ligates a stem-loop adapter to mature tRNAs but not to precursor tRNAs or tRNA fragments. Subsequent TaqMan qRT-PCR amplifies only unmodified regions of the tRNA-adapter ligation products; therefore, FL-PCR quantification is not influenced by tRNA post-transcriptional modifications. FL-PCR has broad applicability for the quantification of various tRNAs in different cell types, and thus provides a much-needed simple method for analyzing tRNA abundance and heterogeneity.
authors.affiliation: a Computational Medicine Center ; Sidney Kimmel Medical College ; Thomas Jefferson University ; Philadelphia , PA USA.
authors.forename: Shozo
authors.initials: S
authors.lastname: Honda
authors.forename: Megumi
authors.initials: M
authors.lastname: Shigematsu
authors.forename: Keisuke
authors.initials: K
authors.lastname: Morichika
authors.forename: Aristeidis G
authors.initials: AG
authors.lastname: Telonis
authors.forename: Yohei
authors.initials: Y
authors.lastname: Kirino
mesh_terms: D001483:Base Sequence; D045744:Cell Line, Tumor; D006801:Humans; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D012343:RNA, Transfer / Q000737:chemistry* / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*
chemical_list: D012343:RNA, Transfer
keywords: FL-PCR; RNA; T4 RNA Ligase 2; TaqMan PCR; tRNA; tRNA abundance; tRNA heterogeneity
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