Qualitative assessment of DNA isolation from fresh, frozen, and ancient human bone using a homogenizer-assisted...

Effective DNA recovery from bone material is essential for applications in biomedical research, clinical diagnostics, and forensic and archaeogenetic investigations. In this study, DNA isolation performance was evaluated in an exploratory manner in human bone samples representing three preservation states: fresh, cryopreserved, and ancient. All samples were processed using a unified bead mill homogenization and magnetic bead-based extraction workflow in order to maintain procedural consistency. DNA quantity and purity were evaluated by spectrophotometry, and amplifiability was assessed using nuclear and mitochondrial PCR assays as well as representative STR profiling. Fresh and cryopreserved samples yielded higher DNA concentrations and more consistent amplification than ancient specimens, in which recovery was primarily constrained by postmortem degradation. PCR success demonstrated a clear dependence on amplicon length, with shorter mitochondrial and nuclear targets amplifying more reliably across all sample types. Due to the limited sample size and the use of a single individual per preservation group, the results are presented as qualitative observations rather than as statistically generalizable conclusions. Within these constraints, the study demonstrates the feasibility of using a standardized mechanical disruption and extraction workflow across bone samples of differing preservation status and provides a methodological reference for future larger-scale studies involving both modern and degraded skeletal material.