title: [Evaluation of 3 methods of DNA extraction from paraffin-embedded material for the amplification of genomic DNA using PCR].
abstract: There are several protocols reported in the literature for the extraction of genomic DNA from formalin-fixed paraffin-embedded samples. Genomic DNA is utilized in molecular analyses, including PCR. This study compares three different methods for the extraction of genomic DNA from formalin-fixed paraffin-embedded (inflammatory fibrous hyperplasia) and non-formalin-fixed (normal oral mucosa) samples: phenol with enzymatic digestion, and silica with and without enzymatic digestion. The amplification of DNA by means of the PCR technique was carried out with primers for the exon 7 of human keratin type 14. Amplicons were analyzed by means of electrophoresis in an 8% polyacrylamide gel with 5% glycerol, followed by silver-staining visualization. The phenol/enzymatic digestion and the silica/enzymatic digestion methods provided amplicons from both tissue samples. The method described is a potential aid in the establishment of the histopathologic diagnosis and in retrospective studies with archival paraffin-embedded samples.
authors.affiliation: Disciplina de Patologia Bucal-Faculdade de Odontologia, USP.
authors.forename: R A
authors.initials: RA
authors.lastname: Mesquita
authors.forename: E K
authors.initials: EK
authors.lastname: Anzai
authors.forename: R N
authors.initials: RN
authors.lastname: Oliveira
authors.forename: F D
authors.initials: FD
authors.lastname: Nunes
mesh_terms: D004247:DNA / Q000302:isolation & purification*; D006801:Humans; D021141:Nucleic Acid Amplification Techniques; D016612:Paraffin Embedding; D016133:Polymerase Chain Reaction / Q000379:methods*
chemical_list: D004247:DNA
vernacular_title: Avaliação de três métodos de extração de DNA de material parafinado para amplificação de DNA genômico pela técnica da PCR.