title: GSM3330275: Female_2; Mus musculus; RNA-Seq
library_construction_protocol: Mouse embryonic fibroblasts (MEFs) from E14 male and female mice (C57BL/6) were generated using standard methods and then were grown in Dulbecco's modified Eagle medium (DMEM, Nacalai Tesque). Media were supplemented with 10% (v/v) fetal bovine serum (FBS) (Gibco), 1% of penicillin-streptomycin (Sigma) at 37°C with 5% CO2. In this study, we used MEFs within 3 passages to avoid replicative senescence. Total RNA was extracted from MEFs using ISOGEN II reagent (Nippon Gene). 500 ng of total RNA were ribosomal RNA-depleted using NEBNext rRNA Depletion Kit (New England Biolabs), and converted to Illumina sequencing library using NEBNext Ultra Directional RNA Library Prep Kit (New England Biolabs). RNA libraries were prepared for sequencing using standard Illumina protocols