miRNA-Seq of Macaca mulatta: adult male PBMC
Small RNA libraries were generated from total RNA as starting material using QIAseq miRNA Library kit (Qiagen, Valencia CA) per manufacturers instructions. Total RNA (10 ng) was 3 (1:5 dilution) and 5 adapter (1:2.5) ligated using miRNA specific adapters. Following reverse transcription (adapters 1:5 dilution), cDNA was size selected using QIASeq miRNA NGS beads and PCR amplified for 19 cycles. Following final cleanups, quality and concentrations of the libraries were measured using Bioanalyzer 2100 (Agilent, Santa Clara CA), and size selected to obtain libraries with a target peak 173bp (average range 145-205 bp) using Pippin Prep (Sage Biosciences, Beverly MA). Final peak sizes and concentration of libraries was verified on the Bioanalyzer. Libraries were sequenced on NextSeq 2500 yielding libraries of 10 million reads each of 75 bp sequenced on single-end mode.