Amplification of mitochondrial DNA from archival temporal bone specimens.
Archival temporal bone collections are an invaluable resource for studying the molecular genetics of many types of otopathology. The irreplaceable nature of temporal bone sections makes efficiency of DNA extraction of paramount importance. Several protocols are available for extracting DNA from fresh and preserved tissue. To establish the best protocol for reliably extracting DNA from celloidin-embedded temporal bone sections, a variety of DNA extraction techniques were tested. Using the optimum protocol, mitochondrial DNA fragments ranging in size from less than 100 base pairs to more than 400 base pairs were amplified, and the authenticity of polymerase chain reaction (PCR) products was confirmed through comparative sequence analysis.
Department of Otolaryngology-Head and Neck Surgery, University of Iowa Hospitals and Clinics, Iowa City 52242.
T A
TA
Simpson
R J
RJ
Smith
D000085:Acetates; D019342:Acetic Acid; D001482:Base Composition; D001483:Base Sequence; D002498:Centrifugation; D011232:Chemical Precipitation; D002725:Chloroform; D003101:Collodion; D017931:DNA Primers; D004272:DNA, Mitochondrial / Q000032:analysis / Q000235:genetics* / Q000302:isolation & purification; D004587:Electrophoresis, Agar Gel; D004591:Electrophoresis, Polyacrylamide Gel; D000431:Ethanol; D005784:Gene Amplification; D006801:Humans; D008967:Molecular Biology; D019800:Phenol; D010636:Phenols; D016133:Polymerase Chain Reaction; D012042:Registries; D015203:Reproducibility of Results; D013701:Temporal Bone / Q000378:metabolism*; D016610:Tissue Embedding
D000085:Acetates; D017931:DNA Primers; D004272:DNA, Mitochondrial; D010636:Phenols; D019800:Phenol; D000431:Ethanol; D002725:Chloroform; D003101:Collodion; D019342:Acetic Acid