Whole-Genome Bisulfite Sequencing in Aging Mice

Whole-genome bisulfite sequencing was performed as previously described in Hadad et al.^{42 (link)} in the hippocampi of 3- and 24-month-old animals. In this instance, bisulfite conversion and library construction were performed according to the manufacturer’s protocol (Accel-NGS Methyl-Seq, Swift Biosciences, Ann Arbor, MI). Libraries were sequenced (HiSeq 2500) to an average coverage depth of > 10× across the entire genome. Alignment and quantitation was as previously described by Hadad et al.^{42 (link)} Quantitation at each CG and CH site covered at ≥10 × was then computed, and an average level of methylation across the genome for each mouse was calculated.