28-30 nt Ribo-seq of caenorhabditis elegans: RNAi against unc-54 in skih-2 animals
Size-selected RNA fragments were treated with PNK buffer (NEB) and ligated to a preadenylated 3' adapter. Excess adapter was removed by sequential digestion with 5' deadenylase and RecJ. Reverse transcription was performed, then cDNA was size selected and extracted by Urea-PAGE. Gel-purified cDNA was circularized using CircLigase ssDNA Ligase. Illumina indexes were added by PCR.