GSM8751986: KI_Sham_2_RRBS; Rattus norvegicus; Bisulfite-Seq
Hippocampi were dissected from each animal following euthanasia and immediately flash-frozen on dry ice. Total RNA was subsequently extracted using the Qiagen AllPrep DNA/RNA Mini Kit. The quality and quantity of the extracted DNA were assessed using a Qubit fluorometer and a TapeStation system. Genomic DNA was digested with Msp1 restriction enzyme and purified using phenol:chloroform extraction and ethanol precipitation. Following Msp1 digestion, genomic DNA underwent blunt-end digestion, phosphorylation, and ligation of adapters with methylated cytosines. Ligated fragments, processed for size selection using agarose gel electrophoresis, were bisulfite converted, amplified by PCR, and then cleaned using AMPure XP beads (Beckman Coulter Life Sciences).