tRAM-seq of zebrafish: embryo 5 hpf, BS, rep2
RNA was extracted with Trizol, tRNA isolated by size selection on denaturing polyacrylamide gel, range 50-150 nt. The RNA was end-repaired by alkaline deacylation and T4 PNK treatment, 3' adapter ligated with T4 RNA ligase 2, reverse transcribed with TGIRT. The cDNA was circularized with CircLigase and amplified with KAPA HiFi polymerase, using NEB Next indexed primers.