title: GSM7360752: 12568_ID_Cfa6.66_ins_BioRep3; Canis lupus familiaris; ChIP-Seq
library_construction_protocol: We extracted nuclei from flash frozen brainstem tissue using the Active Motif High sensitivity kit (Active Motif). Briefly, we re-suspended 50mg of flash-frozen tissue in Tissue fixation solution (4.35ml Molecular grade water, 0.5ml Active Motif 10x PBS, and 140ul 37% (v/v) formaldehyde). We rotated the tube for 8 minutes, added 51.5ul of Active Motif Stop Solution, and followed by 5 minutes of rotation. ChIP-seq libraries were prepared using the Active Motif High sensitivity kit  complemented with the Active Motif Next Gen DNA library Prep kit and Active Motif Next Gen Indexing kit with 15 PCR cycles. 1 ng of DNA was used as starting material for input and ip samples. Post amplification libraries were size selected at 250-450bp in length using Agencourt AMPure XP beads from Beckman Coulter. Libraries were validated using the Agilent High Sensitivity DNA Kit.