Novaseq 6000 s4 sequencers

Total RNA samples were collected and analyzed in the previous study (Gupta et al., 2022 (link)). Briefly, RNA samples were isolated from hPSCs (day 0), hPSC-derived NPCs (day 8), and organoids on days 21, 35, and 49 of differentiation. RNA integrity was assessed (RNA Nano 6000 Assay Kit, Bioanalyzer 2100, Agilent Technologies). Total RNA (400 ng) was used for library preparation using the NEBNext Ultra II RNA Library Prep Kit for Illumina (New England BioLabs, #E7775). Libraries were quantified using Qubit for mass concentration, LabChip for fragments distribution, and qPCR for molar concentration. The qualified RNA-seq libraries were run on NovaSeq 6000 S4 sequencers (Illumina). The MDR1 and OCT2 expression were extracted from the total bulk-RNA seq data and were also compared with developing human kidney datasets (GSE66302). Significant differences between samples were modeled by ANOVA.

DNA samples were diluted in TE buffer and loaded onto Twin-Tec sequencing plate (Eppendorf™) for quality assessment, library preparation, and sequencing from Centre d’expertise et de services Génome Québec (CESGQ). The shotgun PCR-free genomic library was prepared using dual-index adapters from Integrative DNA technologies™. Illumina™ NovaSeq 6000 S4 sequencers provided at least 70 million paired-end reads (150bp) coverage for each sample.