title: Ion Torrent S5 XL sequencing; RNA-sequencing of human umbilical cord-derived MSC (UCX) in normoxia and hypoxia
design: RNA-sequencing of human umbilical cord-derived MSC (UCX) in normoxia and hypoxia
library_construction_protocol: Samples were collected as described in: Santos J, Soares R, Martins JP, Basto V, Coelho M, Cruz P, Cruz H. Isolation method of precursor cells from human umbilical cord. Portugal: Medinfar, ECBio; 2008. (INPI ed., vol. 103843).  Total RNA from UCM-MSC submitted to normoxia or hypoxia for 24h was isolated using the RNeasy Plus Mini Kit (QIAGEN). Ion AmpliSeqâ„¢ Transcriptome Human Gene Expression Kit. Amplicons were digested with the proprietary FuPa enzyme, then barcoded adapters were ligated onto the target amplicons. The library amplicons were bound to magnetic beads, and residual reaction components were washed off. Libraries were amplified, repurified and individually quantified using Agilent TapeStation High Sensitivity tape. Individual libraries were diluted to a final concentration of 50 pM and pooled equally, with twelve individual samples per pool for further processing.