RNA-Seq of Gallus gallus: trachea
RNA samples were isolated from tracheas using RNeasy Fibrous Tissue mini kit (QIAGEN) following the manufacturers instructions. Quantity and quality of isolated RNA samples were checked respectively using Qubit RNA HS assay kit (ThermoFisher Scientific) and RNA 6000 Nano kit using an Agilent 2100 Bioanalyzer (Agilent Technology, Santa Clara, CA). All the samples showed the RNA integrity number (RIN) as > 8. The transcriptome library of each sample was prepared starting from 500 ng of total RNA using the Truseq Stranded mRNA library preparation kit (Illumina Inc., San Diego, CA) following the companys protocols. Each library was run on an Agilent 2100 Bioanalyzer using Agilent High Sensitivity DNA kit to ensure the proper range of cDNA length distribution