Illumina HiSeq 2000 sequencing
Ligation-based small RNA-seq library from Universal Human Reference RNA small RNA fraction (<50 nt); comparison dataset for SEQIRA evaluation.
Small RNA sequencing library prepared from 10 ng DNase I-treated Universal Human Reference RNA (UHRR) small RNA using the ligation-based NEBNext Small RNA Library Prep Kit (New England Biolabs), following the manufacturer's protocol. NEBNext libraries were amplified for 15 PCR cycles, purified with AMPure XP magnetic beads to remove residual primers, multiplexed, and sequenced on an Illumina HiSeq 2000 platform using single-end 50-cycle reads.