degradome-Seq of Humulus lupulus: Wye Target control library
For degradome or PARE sequencing, approximately 150 ng of poly(A)+ RNA was used as input and annealed with biotinylated random primers. RNA fragments were captured using streptavidin beads via the biotinylated random primers. A 5' adapter was ligated exclusively to RNA fragments containing 5'-monophosphates. Reverse transcription was performed, followed by PCR amplification. Libraries were sequenced using only the 5' adapter, capturing the first 36 nucleotides of the inserts, which represent the 5' ends of the original RNAs. Single-end 50 bp sequencing was performed on an Illumina platform.