Protocol for genome-wide differential methylation using long-read DNA sequencing in the cortex of a mouse model of Alzheimer’s disease
Abstract
Third-generation sequencing enables direct detection of genome-wide 5-methylcytosine without bisulfite conversion. Here, we present a protocol for generating long-read DNA sequencing from mouse cortex and performing genome-wide differential methylation analysis. We describe steps for high-molecular-weight DNA extraction, data generation, and downstream analysis. We demonstrate the workflow in the App NL-G-F mouse model of amyloid pathology and provide a reproducible pipeline that links modified-base BAM (modBAM) files to annotated differentially methylated regions and functional enrichment results.
// Source
Authors: Ümran Yaman, Emil Gustavsson, Jasmaine Lee, Hermione Ugolini, Hannah L. Macpherson, Paige Mumford, Frances Wiseman, John Hardy, Dervis A. M. Salih
Institutions: University College London, University of Cambridge, Istanbul University, Istanbul University-Cerrahpaşa, UK Dementia Research Institute, UCL Biomedical Research Centre, UCL Queen Square Institute of Neurology, NIHR Great Ormond Street Hospital Biomedical Research Centre