Characterization of DNA extracted from archival celloidin-embedded human temporal bone sections.
The focus of the present investigation was to study, via molecular biology techniques, the character of the DNA present in individual archival celloidin-embedded human temporal bone sections. Polymerase chain reaction (PCR) amplification of 92 base pair (bp), 121 bp, and 471 bp regions of mitochondrial DNA (mtDNA) extracted from a single archival celloidin-embedded human temporal bone section was used to assess the length of the template DNA extracted. The effects of digestion time and sample motion during the extraction method on DNA concentration was also studied. These data are crucial to determine the limits of applying PCR technology to amplify specific genomic DNA targets located within the human inner ear. Further development of these methods will allow additional molecular temporal bone pathologic studies to be completed and, more specifically, hypotheses regarding the molecular 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 could be tested. The results described should be of great value to those investigators extracting DNA from archival individual human temporal bone sections for PCR assays of specific genetic alterations or infectious agents associated with temporal bone pathologies.
Wackym
P A
PA
Department of Surgery, UCLA School of Medicine 90024-1794, USA.
Chen
C T
CT
Kerner
M M
MM
Bell
T S
TS
D001483:Base Sequence; D017931:DNA Primers; D004272:DNA, Mitochondrial / Q000235:genetics*; D005784:Gene Amplification; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger; D013701:Temporal Bone; D016610:Tissue Embedding
D017931:DNA Primers; D004272:DNA, Mitochondrial; D012333:RNA, Messenger