title: RNA-seq of pig: Piglet longissimus dorsi muscle
design: Total RNA was extracted from the longissimus dorsi muscle of swine using TRIzol reagent (Invitrogen, USA). Each sample was replicated with three biological replicates. After agarose gel electrophoresis and NanoDrop ND-1000 quality inspection and quantification, 1-2 g of total RNA was selected for each sample. mRNA was enriched using the NEBNext  Poly (A) mRNA Magnetic Isolation Module (NEB, USA) to remove rRNA. The RNA sequencing Library was prepared using the KAPA Stranded RNA-Seq Library Prep Kit (Illumina, USA). The strand-specific library construction method based on dUTP was employed to obtain the final RNA sequencing library after PCR amplification. Subsequently, the library's quality was assessed using the Agilent 2100 Bioanalyzer, and the library was quantified using Q-PCR. The pooled libraries from different samples were subjected to high-throughput sequencing using an Illumina NovaSeq 6000 sequencer.