The total RNA was isolated from 4 chicken spleens per treatment group (32 birds total) with the RNAqueous kit (Ambion) using all manufacturers recommendations as previously described (Cheeseman et al. 2007), and treated with DNA-free kit (Ambion). Only samples with an RNA Integrity Number (RIN) greater than 9 were used for cDNA library construction using the TruSeq RNA Library preparation kit v2 (Illumina) that preferentially amplifies PolyA mRNA. The cDNA libraries created from the spleen of 32 chickens were run on the HiSeq 2500 machine (Illumina) with the option of 100 bp single end reads at the Iowa State University DNA Facility. Four lanes were used for RNA-sequencing with 8 multiplexed samples per lane (one sample per each treatment). Individually barcoded samples with 8 different bar codes per lane for multiplexing.
GEO Accession
GSM2266891