title: Illumina HiSeq 4000 sequencing; Gene expression profiling by RNA-seq of human iPSC and iPSC-derived cardiomyocytes from an Yoruban individual (NA19101)
design: Gene expression profiling by RNA-seq of human iPSC and iPSC-derived cardiomyocytes from an Yoruban individual (NA19101)
library_construction_protocol: We used the iPSC line NA19101 (https://www.coriell.org/0/Sections/Search/Sample_Detail.aspx?Ref=GM19101&product=CC) to generate cardiomyocytes. Cardiomyocyte differentiations were carried out similar to the protocol of Burridge et al, 2013 with slight modifications. Briefly, we initiated mesoderm differentiation of the iPSCs with the GSK-3 inhibitor CHIR-99021 trihydrochloride for 24 hours (Tocris #4953). Two days later, cardiomyocyte differentiation was initiated with 2μM Wnt-C59 (Tocris #5148) for 48 hours. Cells exhibited spontaneous beating by day 8-10 and were subsequently selected with lactate on days 15-20 at which time they were harvested and analyzed with flow cytometry for cardiac troponin T. Total RNA was extracted from flash-frozen pellets of 1 million cells using TRI Reagent (Sigma #T9424) and a homogenizer followed by RNA isolation and clean-up using the Direct-zol RNA Kit (Zymo Research #11-331).  RNA-seq libraries were generated with the Illumina TruSeq V2 kit (Illumina, RS-122-2001) and 1μg of RNA, following manufacturer's instructions.