title: chicken_esophagus_3
design: Freshly dissected tissue samples were stored in RNA-later (Sigma-Aldrich), stored at 4 C overnight, and at 80 C afterwards. Samples were then placed in Trizol (VWR) and homogenized prior to RNA extraction according to published protocols (Chomczynski 1993). RNA sequencing for four chicken esophagus samples was performed based on a published approach (Carron et al. 2024). Briefly, sequencing libraries from total RNA of the samples were prepared using the NEBNext Poly(A) mRNA Magnetic Isolation Module and the NEBNext UltraExpress RNA Library Prep Kit for Illumina with Unique Dual Indices (UDIs) according to manufacturer's protocols (New England Biolabs). Libraries were QC-checked on a Bioanalyzer 2100 (Agilent) using a High Sensitivity DNA Kit for correct insert size and quantified using Qubit dsDNA HS Assay (Invitrogen). Libraries were sequenced on a P4 flowcell of a NextSeq2000 instrument in 1x50bp single end sequencing mode. On average, 22 million reads per sample were generated. Reads in fastq format were generated using the Illumina bcl2fastq command line tool (Illumina, v2.19.1.403).