NO_lung_1 (CRX1865409; CRA028061)
Total RNA was extracted from samples using the chloroform method. The concentration and purity of the total RNA were measured using the Agilent Bioanalyzer 2100 System. Subsequently, RNA integrity was assessed using an Agilent Bioanalyzer. RNA samples exhibiting good integrity and acceptable purity were selected for cDNA library construction. The specific workflow included: oligo-dT bead enrichment of poly(A)-tailed mRNA, mRNA fragmentation, first-strand and second-strand cDNA synthesis, end repair, A-tailing, adapter ligation, and library amplification. The final libraries were sequenced on the Illumina platform. Raw sequencing data quality was assessed using FastQC (v0.11.9). Subsequently, quality control of the raw reads was performed using SeqPrep (v1.3.2) to obtain clean reads. The clean reads were aligned to the chicken (Gallus gallus) reference genome (GRCg7b) using HISAT2 (v2.2.1). Transcripts were assembled using StringTie (v2.1.7), and gene expression quantification was performed using the featureCounts tool within the subread software package (v2.0.3) to generate raw gene expression count data submitted_instrument_model: Illumina HiSeq X Ten