tRNAs modified with multiple methylations (m1A, m3C, m1G, m2,2G) can be simultaneously and quantitatively profiled at single-base resolution by tRNA Modification Seq - m1A/m3C/m1G/m2,2G. These modifications disrupt Watson–Crick base pairing and can induce mismatch mutations due to misincorporation by reverse transcription.
To identify modification sites and quantify the methylation in a tRNA, the highly-optimized tRNA-seq is performed on the tRNA with and without demethylase treatment. The modification is detected by methylation induced mutation index (MI) at each base position compared with its demethylase-treated tRNA. Among m1A, m3C, m1G, and m2,2G modifications, the m1G and m2,2G are further distinguished by their known distinct base positions in mammalian tRNAs (m1G at 9 and 37; m2,2G at 26 tRNA base positions).
Benefits:
• Multiple methylation modification detection. Simultaneously profile m1A, m3C, m1G and m2,2G modifications in tRNAs
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High efficiency methylation removal. Efficiently remove internal and terminal modifications in the demethylase treated tRNA. Learn more>>
• Single-base resolution. Identify the exact sites of m1A, m3C, m1G and m2,2G methylation modifications by comparing demethylase-untreated tRNA with demethylase-treated tRNA side by side.
• Comprehensive tRNA Epitranscriptomic Analysis. Analyze modifications sites, methylation levels, and tRNA expression all at once.
• High performance tRNA Sequencing. Highly optimized tRNA-seq with rigorous QC process, overcoming difficulties of regular sequencing for tRNA.
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Figure 1. Workflow of tRNA Modification Seq. m1A, m3C, m1G, and m2,2G modifications are identified by their induced misincorporation mutations in reverse transcription compared with demethylase-treated tRNA.