GSM5243495: TT-249_PHA-767491; Homo sapiens; RNA-Seq
RNA Extraction: Approximately 10^5 to 10^6 iPSC-CMs were lysed with TRI Reagent (Zymo Research) and frozen at -80 °C to ensure complete cell lysis. Total RNA was extracted and washed from lysed cells using the Direct-zol-96 RNA Kit (Zymo Research) according to the manufacturer's instructions. Samples were treated with DNase I for 15 minutes at room temperature. cDNA was reverse transcribed from 1 μg of RNA through random hexamers using the SuperScript-III kit (Invitrogen) according to the manufacturer's instructions. Real-time qPCR reactions were performed using the TaqMan universal PCR master mix (Applied Biosystems) with the TaqMan probes listed in Table S5 (Life Technologies). Library Preparation: 100ng total RNA was extracted via polyA-tail specific protocol according to Illumina protocol. RNA library preparation was achieved by using TruSeq Stranded mRNA kit (Illumina) which also removes ribosomal RNA. The libraries were sequenced as 100bp single-end reads using Illumina NovaSeq SP with an average of 30M reads per sample.
GEO Accession
GSM5243495