GSM9192096: Atf3 KO Mes cells day 0 rep 2; Mus musculus; RNA-Seq
Total RNA was extracted from mESCs at days 0, 3, 6, and 9 of differentiation using TRIzol reagent (Invitrogen) according to the manufacturer's instructions. RNA integrity was assessed using an Agilent 2100 Bioanalyzer. RNA-seq libraries were prepared using the NEBNext Ultra II RNA Library Prep Kit for Illumina (New England Biolabs) following the manufacturer's protocol. Poly(A)+ RNA was purified from total RNA using oligo(dT) beads, fragmented to ~200 bp, and reverse transcribed to synthesize first-strand cDNA. Second-strand cDNA was generated, followed by end repair, A-tailing, and ligation of Illumina sequencing adapters. Libraries were PCR-amplified (12 cycles) and sequenced on an Illumina NovaSeq 6000 platform to generate 150 bp paired-end reads.