tRNAs modified with multiple methylations (m1A, m3C, m1G, m2,2G) can be simultaneously and quantitatively profiled at single-base resolution by m1A/m3C/m1G/m2,2G tRNA-seq. These modifications disrupt Watson–Crick base pairing and can induce mismatch mutations due to misincorporation by reverse transcription. To identify their modification sites and quantify the methylation, highly-optimized tRNA-seq is performed on the tRNA sample with and without demethylation treatment. The modification is detected by methylation induced mutation index (MI) at each base position compared with demethylated tRNA "input" reference. Among m1A, m3C, m1G, and m2,2G modifications, m1G and m2,2G are further distinguished by their known distinct locations in cytoplasmic and mitochondrial tRNAs in mammals (m1G at tRNA base positions 9 and 37; m2,2G at 26).
Benefits:
• Multiple methylation modification detection. Simultaneously profile m1A, m3C, m1G and m2,2G modifications in tRNAs
• Single-base resolution. Identify the exact sites of m1A, m3C, m1G and m2,2G methylation modifications by comparing demethylation untreated sample with demethylation treated reference side by side.
•
High efficiency methylation removal. Efficiently remove internal and terminal modifications in the demethylation treated reference. Learn more>>• 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 m1A/m3C/m1G/m2,2G tRNA-Seq. m1A, m3C, m1G, and m2,2G modifications are identified by their induced misincorporation mutations in reverse transcription compared with demethylation treated tRNA-seq reference.