Differentially methylated Embryonal Fyn-associated Substrate (EFS) gene as a blood-specific epigenetic marker and its potential application in forensic casework

Highlights • A novel blood-specific differentially methylated locus (EFS) was investigated. • Most CpGs, especially CpG4, was found highly methylated in whole blood. • EFS methylation has the potential to differentiate between whole and menstrual blood. • The designed bisulfite pyrosequencing assay is both accurate and reproducible. • The method’s applicability to casework was demonstrated via aged and mixed stains.