Linseed diet replicate 3
Total RNA was isolated from Milk Somatic Cells (MSC). MSC were pelleted from 250 mL fresh milk at 110010 min at 4 C. The cell pellet was washed twice in phosphate-buffered saline solu-tion, pH 7.2 with 0.5 mM EDTA and resuspended in 3 ml of TriZol reagent. RNA extraction continued following the TriZol protocol and precipitation in isopropanol was car-ried on at 80 C for 30 min. Then, a DNAse I (Roche, Basel, Switzerland) treatment wasperformed to completely remove genomicDNA contamination. Poly-A RNAs were isolated from totalRNA and chemically fragmented. First and second strand cDNA synthesis were followedby end-repair and adenosines were added to the 3 ends. Adapters were ligated to thecDNA and 200  25 bp fragments were gel purified and enriched by PCR. Overall 8 librar-ies were quantified using Bioanalyzer 2100 (Agilent Technologies, Santa Clara, CA) andrun on the Illumina HiSeq2000 (Illumina Inc.) using version 3 reagents. Paired-read se-quences, 125 nt in length, were collected.