GSM4890451: inhibitor-3; Homo sapiens; ncRNA-Seq
Total RNA isolation, including DNase digestion, was performed using the miRNeasy Mini Kit (Qiagen, #217004, Chadstone, Australia) and RNase-free DNase Set (Qiagen, #79254, Chadstone, Australia) as instructed by the manufacturer. Quantification of the final RNA concentration was performed via UV spectrophotometry (NanoDrop™ 2000 Spectrophotometer, Thermo Fisher Scientific, Wilmington, USA). Libraries of small RNAs were prepared from 150 ng of RNA using the NEBNext™ kit (New England Biolabs). After reverse transcription, 15 cycles of PCR were performed to enrich for successfully ligated molecules. The libraries were size selected using 3 % agarose gels targeting a 122–182 bp range. Size-selected libraries were run on Bioanalyzer high-sensitivity DNA chips to confirm size, minimal adapter dimers, and to estimate yield.