[Evaluation of 3 methods of DNA extraction from paraffin-embedded material for the amplification of genomic DNA using PCR].
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.
Mesquita
R A
RA
Disciplina de Patologia Bucal-Faculdade de Odontologia, USP.
Anzai
E K
EK
Oliveira
R N
RN
Nunes
F D
FD
D004247:DNA / Q000302:isolation & purification*; D006801:Humans; D021141:Nucleic Acid Amplification Techniques; D016612:Paraffin Embedding; D016133:Polymerase Chain Reaction / Q000379:methods*
D004247:DNA
Avaliação de três métodos de extração de DNA de material parafinado para amplificação de DNA genômico pela técnica da PCR.