GSM4879289: BJFF_d28; Homo sapiens; RNA-Seq
To determine transcriptome profiles over the course of differentiation, three unique hiPSCs lines (ATCC, BJFF, and STAN) as biological replicates at various differentiation stages (6 mesodermal and 5 chondrogenic stages per cell line; i.e., total 33 samples) were collected for bulk RNA-seq. Cell samples were thawed on ice, and pellet samples were homogenized with zirconia beads (BioSpec Products, # 11079110zx) and a miniature bead beater. RNA was then isolated from all samples using the Total RNA Purification Kit according to the manufacture's protocol (Norgen Biotek, #37500). RNA was eluted in 20 µl of diethylpyrocarbonate-treated water. The quality and quantity of RNA from each sample was evaluated by RNA Analysis ScreenTape (Agilent, #5067-5576) on bioanalyzer (Agilent 4200 Tapestation). Only samples with a RIN value larger than 0.8 were submitted to the Genome Technology Access Center (GTAC sequencing core) at Washington University in St. Louis for library preparation and bulk RNA-seq. Libraries were prepared using TruSeq Stranded Total RNA with Ribo-Zero Gold kit (Illumina). Sequencing was performed on a Illumina HiSeq 2500 (1 x 50 bp reads) with a sequencing depth of 30 million reads per sample. RNA libraries were prepared for sequencing using standard Illumina protocols Bulk RNA-seq