title: GSM4704322: undifferentiated RNA-Seq3; Homo sapiens; RNA-Seq
library_construction_protocol: For each sample, 1µg of high quality (RNA integrity number = 10 for all samples) total RNA was subjected to library preparation for mRNA sequencing using the TruSeq Stranded mRNA Kit (Illumina), as per the manufacturer's instructions. Briefly, mRNA was denatured (65°C, 5 min) and ligated to oligo-dT magnetic beads (room temperature, 5 min), after which unbound RNA and non-specifically bound rRNA was discarded. mRNA was then heat fragmented (94°C, 8 min), following which reverse transcription and adapter ligation were performed as per the manufacturer's instructions. Enrichment of cDNA fragments was subsequently conducted via 15 cycles of PCR, after which libraries were normalised, pooled and subjected to 151 cycles of sequencing using the NextSeq 500 system. Total RNA was extracted from all groups by harvesting cells in 1mL TRIzol reagent (ThermoFisher, Waltham, MA, USA), as per the manufacturer's instructions, with RNA precipitated at –30°C overnight to increase yield.
attributes.tag: GEO Accession
attributes.value: GSM4704322