Amplification of RNA from archival human temporal bone sections

Background The amplification of DNA from celloidin-embedded human temporal bone sections by polymerase chain reaction (PCR) has been applied to some a uditory diseases, such as herpes zoster oticus and hearing loss caused by t he mutations of mitochondrial DNA However, few studies have reported detect ion of RNA from temporal bone sections. Objectives: Because RNA analysis fr om temporal bone sections may elucidate the development of the diseases in the auditory, vestibular, and facial nerves, the authors investigated wheth er RNA in these sections can be amplified by reverse transcription (RT)-PCR Methods: Sections that were formallin-fixed, decalcified, and embedded bet ween 1972 and 1986 were used. Nucleic acid was extracted from the celloidin -embedded temporal bone sections and subjected to RT-PCR, Human alpha-tubul in RNA was reverse transcribed to cDNA and amplified by nested PCR using tw o sets of primers that were designed to distinguish cDNA from genomic DNA b ased on the presence of an intron between the primers. Results: Human alpha -tubulin RNA was detected in 11 of 14 temporal bone sections (79%) by RT-PC R RNA was detected in even the oldest sections, which were processed in 197 2, Conclusions: These results indicate that RNA can be analyzed from archiv al celloidin-embedded human temporal bone sections.