GSM5942373: C_3; Homo sapiens; RNA-Seq
Total RNA were isolated from undifferentiated and differentiated isogenic control and R723C hiPSCs using the RNeasy kit (Qiagen) according to the manufacturer's protocol. Briefly, washed the cells once with sterile PBS and lysed in RLT-lysis buffer for specific time points. RNA isolation was performed as per the protocol, followed by on-column DNA digestion to remove any traces of DNA as per the instructions. For cDNA synthesis, 100-1000 ng of total RNA was used and synthesized using the SuperScript IV VILO kit (Thermo Fisher Scientific) according to the manufacturer's protocol. Quantitative PCR (qPCR) was performed using gene-specific oligos and the SYBR-green method. The total RNA was isolated as described above. For the bulk-RNAseq analysis, 1 μg of total RNA was submitted to Novogene (https://en.novogene.com/) for sequencing and data analysis. At Novogene, cDNA libraries are sequenced using the state-of-the-art Illumina NovaSeq platforms, which utilize a paired-end 150 bp sequencing strategy (short-reads).