Small RNA-seq (miRNA-seq) of primary human cells
RNA was isolated using the miRNeasy kit from Qiagen according to the manufacturers protocol. RNA integrity was assessed using Agilent BioAnalyzer and the RNA concentrations were measured using a NanoDrop 2000 UV-Vis Spectrophotometer. Small RNA libraries were prepared using the Illumina TruSeq Small RNA Library Preparation kit according to the manufacturers protocol, but purified using a Pippin Prep with a 3% Agarose Gel Cassette (Sage Science) and a size selection of 122-157bp. Multiplexed sequencing was performed as single read 50 base pair, using rapid run mode, and v2 chemistry on a HiSeq2500 system.