Comparative tRNA-seq for rapid profiling of tRNA modifications in a non-model organism

tRNAs are heavily decorated with post-transcriptional modifications (tRNA modifications). Profile of tRNA modifications in non-model organisms are largely uncharacterized. Here using high-throughput sequencing, sites and frequency of tRNA modifications are predicted in Vibrio cholerae and Escherichia coli. During cDNA synthesis, some modifications cause misincorporation of a wrong base or termination of reverse transcription (RT). Using these RT-derived signatures we aim to explore organism-specific modifications and to track changes in modification frequency among different cellular conditions. Overall design: High throughput sequencing of tRNA fractions from Escherichia coli (log and stationary phase) and Vibrio cholerae (log phase, stationary phase, and cecal fluid from infant rabbit) was performed using Illumina miSeq.