tRNA-seq
The tRNA fraction was isolated using AX-20 columns (Macherey-Nagel). The tRNA was deacylated and dephosphorylated and 165 ng tRNA was used to ligate library-specific adapters. Reverse transcription was performed using either Induro (New England Biolabs) or SuperScript IV (Thermo Fisher Scientific). RNA was removed by addition of 5 M NaOH. Gel-purified cDNAs were circularized and amplified using Q5 Polymerase (New England Biolabs). Libraries were resolved on a 6% PAGE gel, and 130-230 bp (mim-tRNAseq) or 140-260 bp (QuantM-tRNAseq) fragments were excised and eluted. The concentration was estimated using the NEBNext Library Quant Kit for Illumina (New England Biolabs).