Epigenetic landscape of hormone-independent sexual dimorphism and characterization of canine XIST - WGBS
Eight beagle dogs (four males and four females) were investigated for sexual dimorphism in their transcriptomes. All dogs were five-year-old and had been neutered (castrated or spayed) before one year of age, and their healthy physical conditions were maintained through continuous monitoring, vaccination, and anthelmintic treatment. The experiment was ethically approved by the Institutional Animal Care and Use Committee of National Institute of Animal Science (NIAS 2022-586). Whole blood samples were collected using a 21-gauge needle and 5-mL syringe from the jugular vein of each dog. DNA was extracted from whole blood using an Exgene Blood SV mini kit (GeneAll Biotechnology, Seoul, Republic of Korea) according to the manufacturerâ€™s protocol.  Libraries were prepared using the xGen Methyl-Seq Library Prep Kit (Integrated DNA Technologies, Coralville, IA, USA). The input DNA concentration was analyzed using Qubit. A total of 100 ng genomic DNA was fragmented to an estimated size of 350 bp using the S2 Focused Ultrasonicator system (Covaris, Woburn, MA, USA). 