title: Stranded RNA-seq of Bos taurus: Dominette subcutaneous fat
design: The transcriptomes of 23 tissues from the Hereford cow, L1 Dominette 01449 (the reference genome individual), and testis from SuperBull 99375 were sequenced by the Meat Animal Research Center (MARC, USDA, Clay Center, NE) using the Illumina NextSeq500. Total RNA of each tissue was extracted using Trizol reagent (ThermoFisher Scientific) and the protocol included. Briefly, approximately 100 mg of tissue was ground in a mortar and pestle cooled with liquid nitrogen, and the powder was transferred to a tube with 1 ml of Trizol reagent added and mixed by vortexing. After 5 minutes at room temperature, 0.2 mL of chloroform was added and the mixture was shaken for 15 seconds and left to stand another 3 minutes. The tube was centrifuged at 12,000 x g for 15 minutes at 4ĄC. The RNA was precipitated from the aqueous phase with 0.5 mL of isopropanol. The RNA was further purified with extended DNase digestion to remove potential DNA contamination. The RNA quality was assessed with Fragment Analyzer (AATI, IA). Only RNA samples of RQN above 7.0 were used for library construction. Short read RNAseq libraries were prepared using TruSeq stranded mRNA LT kits and supplied protocol (Illumina, CA), and sequenced on the NextSeq500 platform using 2x75 paired end reads and v2 sequencing chemistry.