title: GSM6661413: IMR32_DMSO_Rep4 (RIBOseq); Homo sapiens; OTHER
library_construction_protocol: 15µg of the lysates were treated with 7,5 units RNase I, purified with the Microspin S-400 columns and the Zymo RNA clean and concentrator kit (Zymo research, ZY-R1015). The eluted samples were size selected (28-30 nt) on a 15% PAGE urea gels. Samples were 3' dephosphorylated with T4 PNK followed by library prep using the Small RNA library prep kit (Lexogen, 052.96). rRNA was blocked with LNA probes during the cDNA synthesis step of the library prep using the FastSelect H/M/R kit from Qiagen. Library amplification was done according to Lexogen small RNA kit. The PCR products were purified using Ampure XP beads. Library quality control was done on a DNA high sensitivity chip (BioAnalyzer Agilent, 5067-4626). Library concentrations were measured with qPCR according to Illumina's protocol. Libraries were sequenced on 2 NextSeq500 high output runs with 76bp single reads.