GSE111230
Induced pluripotent stem cells of patients with Tetralogy of Fallot reveal transcriptional alterations in cardiomyocyte differentiation
We generated induced pluripotent stem cells (iPSCs) of two patients with Tetralogy of Fallot (TOF) and three healthy relatives of two unrelated familes. We furhter performed mRNA sequencing of iPSCs (day 0) and derived cardiomyocytes (CMs) at day 15 and 60 of patients and healthy relatives. Overall design: Total RNA was isolated using Direct-zol RNA Kits (Zymo Reasearch) and purified with Dynabeads Oligo(dT) (Thermo Fisher Scientific) to get poly(A) RNA. The RNA libraries were prepared with the ScriptSeq v2 RNA-Seq Library Preparation Kit (Illumina). The libraries were evaluated with the Qubit dsDNA HS Assay Kit (Thermo Fisher Scientific) and the Agilent High Sensitivity DNA Kit (Agilent Technologies). Paired-end sequencing was performed on either HiSeq 2500 (2x51 bp) or NextSeq (2x76 bp) device (Illumina). For the latter, the NextSeq 500/550 v2 kits (Illumina) were used. All kits were applied according to the manufacturer's instructions. Overall, mRNA sequencing was performed for 31 samples of 5 individuals for the following three differentiation stages: pluripotent iPSC (day 0), iPSC-CM (day 15) and CM (day 60). Sequencing was separated into two runs, all day 0 samples and three day 15 samples (Mother TOF-02) were sequenced with Illumina's HiSeq 2500. The remaining samples were sequenced using Illumina's NextSeq.