The identification of genes regulating milk secretion by mammary glands is a key-step for exploiting changes in milk composition induced by different diet regimens. Linseed supplementation is a reliable feeding strategy to enhance polyunsaturated fatty acid content in milk fat from sheep, increasing milk quality and nutraceutical value. To investigate the molecular bases of diet induced differences in milk composition, we collected milk from dairy ewes at 90 day of lactation and after 3 weeks of diet supplementation with extruded linseed. The milk of dairy ewes contains milk somatic cells mostly derived by mammary glands. After isolating milk somatic cells by centrifugation, RNAs were purified from these cells, and Illumina RNA sequencing was performed to analyze RNA synthesis. Our data provide a resource (available at Gene Expression Omnibus database under GSE89163) to be employed for comparative analyses of gene expression in milk somatic cells in different breeds and different diets, with the long-term aim of developing strategies to improve sheep milk quality.