tRNA_seq of Caenorhabditis elegans: Young Adult
the purified tRNA underwent a critical pre-treatment step involving mA and mC demethylation to reduce sequencing biases, and was then partially hydrolyzed using the Hydro-tRNA seq protocol. These processed fragments were re-phosphorylated and converted into sequencing libraries with the NEBNext Multiplex Small RNA Library Prep Set for Illumina. The resulting libraries underwent stringent size selection (~140-155 bp, corresponding to ~19-35 nt tRNA fragments) and were rigorously qualified and quantified on an Agilent 2100 BioAnalyzer prior to being pooled in equimolar amounts for sequencing on an Illumina platform.