A method was developed for fast and efficient isolation of DNA from formalin-fixed, decalcified, celloidin-embedded human temporal bone sections for subsequent use in polymerase chain reaction (PCR) DNA amplification. The method relies on the use of an enzymatic digestion with proteinase K to release and solubilize the patient's DNA from an individual 20- to 25-microns temporal bone section. The method described should be of great value to those investigators extracting DNA from archival individual human temporal bone sections for polymerase chain reaction assays of specific genetic alterations associated with temporal bone pathologies. The molecular characterization of viral infections, oncogenes, or other etiological agents of disease using PCR could provide important information regarding the etiopathogenesis of many auditory, vestibular, and facial nerve disorders, such as autoimmune hearing loss, congenital hearing losses, Meniere's disease, otosclerosis, or Bell's palsy.