The 12 o’clock assay: an optimized dodecaplex droplet digital PCR assay for robust DNA methylation quantification and epigenetic clock-based age-predictions
Year: 2026
Type: article
Abstract: DNA methylation quantification in age-prediction models relies on only a few techniques with inherent limitations and biases, reducing the accuracy and reproducibility. We developed a new droplet digital PCR assay format for accurate DNA methylation quantification, based on three LNA-containing hydrolysis probes to detect methylated and unmethylated alleles. Using a 12-plex assay we simultaneously quantified methylation at six CpGs ( ASPA , C1orf132 , CCDC102B , EDARADD , ELOVL2 , FHL2 ) in 351 blood samples (age range: 0–95 years), showing strong correlations with age (| r| > 0.83). Several age-prediction models were developed, achieving high performance on testing sets ( R 2 : 0.933–0.973, MAE: 3.18–5.18, RMSE: 4.71–7.27).
Source: Clinical Epigenetics
Institutions: Fondation Jean Dausset-CEPH, Autorité de sûreté nucléaire et de radioprotection, Inserm, Institut Curie, Conservatoire National des Arts et Métiers, Génomique, bioinformatique et chimie moléculaire, Université Paris Cité, Institut National de Recherche pour l'Agriculture, l'Alimentation et l'Environnement, Université Sorbonne Paris Nord, Equipe de Recherche en Epidémiologie Nutritionnelle, Centre de Recherche Épidémiologie et Statistique, HIPI - Immunologie humaine, physiopathologie et immunithérapie, Centre Hospitalier Régional et Universitaire de Nancy, Défaillance Cardiovasculaire Aiguë et Chronique, Université de Lorraine, Commissariat à l'Énergie Atomique et aux Énergies Alternatives, Centre National de Recherche en Génomique Humaine
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $3,890
OpenAlex ID: W7140225114