Detection of DNA mutations in acid formalin-fixed paraffin-embedded archival tumor specimens by polymerase chain reaction-single strand conformation polymorphism analysis.
A single strand conformation polymorphism analysis on polymerase chain reaction (PCR)-amplified genomic DNA was used for the detection of DNA mutations in acid formalin-fixed, paraffin-embedded tissues. Fifty non-small cell lung cancers were screened for mutations in the exon 5 of the p53 tumor suppressor gene. Structural abnormalities of the gene were found in nine (18%) of the tumors examined. The results show that paraffin-embedded tissues fixed in unbuffered formalin can be a good source of DNA for a mobility shift analysis by neutral polyacrylamide gel electrophoresis. The described procedure allows retrospective genetic studies on paraffin blocks available in all pathology departments.
Marchetti
A
A
Institute of Pathological Anatomy and Histology, University of Pisa, Italy.
Merlo
G
G
Buttitta
F
F
Pellegrini
S
S
Callahan
R
R
Bistocchi
M
M
Squartini
F
F
D001483:Base Sequence; D002289:Carcinoma, Non-Small-Cell Lung / Q000235:genetics*; D004247:DNA / Q000032:analysis* / Q000187:drug effects; D005404:Fixatives; D005557:Formaldehyde; D016158:Genes, p53 / Q000235:genetics; D006801:Humans; D008175:Lung Neoplasms / Q000235:genetics*; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics*; D016612:Paraffin Embedding / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*; D018807:Polymorphism, Single-Stranded Conformational; D012189:Retrospective Studies; D016707:Tissue Fixation / Q000379:methods
D005404:Fixatives; D005557:Formaldehyde; D004247:DNA