GSM6008419: ADSC, Donor 66, FBS culture condition; Homo sapiens; RNA-Seq
Total RNA was isolated using Trizol reagent (Invitrogen). RNA quality was assessed using Agilent 2100 bioanalyzer with the RNA 6000 Nano Chip (Agilent Technologies, Amstelveen, Netherlands). RNA was quantified using ND-2000 spectrophotometer (Thermo Fisher Scientific). AmpliSeq libraries were constructed and sequenced on the Ion Torrent S5xl platform (Thermo Fisher) according to the manufacturer's instructions. For human gene analysis in cell lines, the Ion AmpliSeq Transcriptome Human Gene Expression Kit is designed for targeted amplification of over 20,000 human RefSeq genes simultaneously in a single primer pool. A short amplicon (~ 110 base pairs (bp)) is amplified for targeted gene. For each sample, 30 ng of total RNA was used for cDNA library preparation. Multiple libraries were multiplexed and clonally amplified using the Ion Chef System (Thermo Fisher).