GSM4041543: 071-2-NoIns-72hrs; Homo sapiens; RNA-Seq
RNA was extracted as described earlier. [DOI:https://doi.org/10.1016/j.stem.2016.11.005]  Briefly:  Cells were washed once with PBS and RNA was extracted using Purelink RNA mini kit (Thermo Fisher Scientific). RNA integrity was evaluated using 2100 Bioanalyzer via the RNA 6000 Nano assay (Agilent) or Fragment Analyzer (Advanced Analytical). Only samples with RQN/RIN values equal or greater than 7.0 were used. library was constructed as described earlier. [DOI:https://doi.org/10.1016/j.stem.2016.11.005]  Briefly:  Library preparation was done using standard TruSeq RNA Sample Prep Kit v2 protocol (Illumina). mRNA was isolated and fragmented. cDNA was generated using random hexamers, end-repaired and ligated with respctive adaptors for sequencing.  AMPure XP beads (Beckman Coulter) was used size selection and purification of the library. During PCR amplification step the illumine recommended  6 bp barcode bases were introduced at one end of the adaptors. Before load onto the sequencer,  the size and concentration of the RNA-seq libraries were measured by Qubit fluorometry (Thermo Fisher Scientific) and Bioanalyzer. Illumina HiSeq 2500 System  was used for sequencing of the mRNA libraries with 100 nucleotide single-end reads, according to the Illumina's standard protocol.