RNA-Seq of chicken:6w male wattles
Total RNA was extracted from wattle samples using TRIzol reagent. RNA integrity was verified by agarose gel electrophoresis and quantified with a NanoDrop spectrophotometer.Sequencing libraries were constructed following the standard protocol of the NEBNext Ultra RNA Library Prep Kit for Illumina. Briefly, mRNA was purified from total RNA using poly-T oligo-attached magnetic beads and then fragmented. First-strand cDNA was synthesized using random hexamer primers, followed by second-strand synthesis. The resulting cDNA fragments were end-repaired, adenylated at the 3' ends, and ligated with Illumina adapters. After purification, the adapter-ligated fragments were amplified by PCR to enrich the final library. Library quality was assessed using an Agilent Bioanalyzer 2100 system.The prepared libraries were sequenced on an Illumina NovaSeq 6000 platform to generate 150 bp paired-end reads.