GSM9486094: WT3 (M1_alpha), RNA-seq_H_E9-5_WT_rep3; Mus musculus; RNA-Seq
Stage-matched embryonic hearts were micro-dissected and collected in RNAlater (Sigma, R0901). Total RNA was extracted using the Qiagen RNeasy Micro Kit (Cat. No. 74004). Per replicate, RNA quantity and integrity were evaluated using a Qubit 4.0 Fluorometer with the Qubit RNA HS Assay Kit (Thermo Fisher Scientific, Q32855) and a Fragment Analyzer System with the Fragment Analyzer RNA Kit (Agilent, DNF-471), respectively. Sequencing libraries were generated from 300 ng of total RNA using the TruSeq Stranded mRNA Library Prep Kit (Illumina, 20020595) in combination with TruSeq RNA UD Indexes (Illumina, 20022371), according to the manufacturer's protocol. Pooled libraries were quantified using a Qubit 4.0 fluorometer with the Qubit dsDNA HS Assay Kit (Fisher Scientific, Q32854) and assessed for fragment size distribution on an Agilent Fragment Analyzer with the HS NGS Fragment Kit (Agilent, DNF-474). Equimolar amounts of each library were then pooled and sequenced in paired-end mode on an Illumina NovaSeq 6000 platform using the NovaSeq 6000 SP Reagent Kit v1.5 (300 cycles; Illumina, 20028400).