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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.

SUBMITTER: Honda S

PROVIDER: S-EPMC4615770 | biostudies-literature | 2015

REPOSITORIES: biostudies-literature

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Four-leaf clover qRT-PCR: A convenient method for selective quantification of mature tRNA.

Honda Shozo S Shigematsu Megumi M Morichika Keisuke K Telonis Aristeidis G AG Kirino Yohei Y

RNA biology 20150101 5

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 un ...[more]

PMID: 25833336

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Four-leaf clover qRT-PCR: A convenient method for selective quantification of mature tRNA.

Publications

Four-leaf clover qRT-PCR: A convenient method for selective quantification of mature tRNA.

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