title: RNA-seq of Meleagris gallopavo: male cecal tonsil
design: RNA-seq was performed to understand acute responses to S. Reading and the effect of vaccination on the gene expression in turkey cecal tonsil. Total RNA was extracted from turkey cecal tonsil tissues collected at 2 days post inoculation from the M/M, M/R, BBSoo/R, and AMEoo/R groups (n = 12 birds/group) using the RNAqueous Total RNA Isolation kit with on-column DNase treatment using the QIAGEN RNase-free DNase Set. Stranded mRNA-seq libraries were constructed using the NEBNext Ultra II Directional RNA Library Prep kit with the NEBNext Multiplex Oligos for Illumina (96 Unique Dual Index Primer Pairs) kit for sample indexing. Libraries were sequenced on a NovaSeq 6000 using 4 lanes (n = 4 technical replicates/sample) of an S4 flow cell to produce 100-cycle paired-end reads.