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
•
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.

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.