GSM5235908: Treatment A rep 4; Gallus gallus; RNA-Seq
On day 46, birds were euthanized and Liver tissue samples were taken, preserved in RNAlater, placed at 4°C for 24 hours then stored at -20°C. RNA was isolated with RNAeasy kit (QIAGEN-Hilden, Germany) according to manufactures protocol from liver samples of both the highest density (Treatment D) and the lowest density (Treatment A). Briefly, tissue samples were initially homogenized in kit lysis buffer and centrifuged. The supernatant was combined with equal volume 70% ethanol and loaded on the provided spin columns. Bound RNA was washed with provided buffers and eluted in nuclease free water. Final RNA sample yields were quantified spectrophotometrically using NanoDrop 2000 (Thermo Fisher Scientific-Waltham, MA). RNA samples were submitted to NC State Universities Genomic Sciences Laboratory for library preparation and sequencing using Illumina HiSeq (Illumina-San Diego, CA). Final RNA sample yields were quantified spectrophotometrically using NanoDrop 2000 (Thermo Fisher Scientific-Waltham, MA). RNA samples were submitted to NC State Universities Genomic Sciences Laboratory for library preparation and sequencing using Illumina HiSeq (Illumina-San Diego, CA). Libraries were prepared according to Illumina's instructions and each library consisted of equal amounts pooled RNA from 4 samples of the same treatment and 4 libraries were prepared for each of the 2 treatments.