GSM9679083: RNA-seq of  CM-specific ATF4 CDS KO mice  E11.5  heart replicate 3; Mus musculus; RNA-Seq
Total RNA was extracted from cultured primary cortical neurons using the Quick-RNA Miniprep Kit (Zymo Research; Cat# R1054) following the manufacturer's instructions. RNA concentration and purity were assessed using a Nanodrop spectrophotometer. For downstream applications, equal amounts of RNA were used for either RNA-seq library preparation or reverse transcription into cDNA using the High-Capacity cDNA Reverse Transcription Kit (Invitrogen™). Gene expression validation was performed by quantitative PCR (qPCR) using the Applied Biosystems 7300 Real-Time PCR System, and analyzed with the ΔΔCt method. RNA-seq libraries were constructed following ribosomal RNA (rRNA) depletion protocols to enrich for non-rRNA transcripts. Total RNA was subjected to rRNA depletion using a commercial RNA depletion kit according to the manufacturer's instructions. Libraries were then generated from the rRNA-depleted RNA and sequenced on an Illumina NovaSeq 6000 platform using paired-end sequencing.