Evaluation of the genomic DNA extracted from formalin-fixed, paraffin-embedded oral samples archived for the past 40-years
BACKGROUND: The most common human archival specimens are formalin-fixed, paraffin-embedded tissues (PETs). DNA can be extracted from PETs, but sometimes, it is unsuitable for molecular techniques as slow degradation of DNA occurs with time. OBJECTIVE: The aim of this study was to verify and discuss...
| Autores: | , , , , |
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| Tipo de recurso: | artículo |
| Estado: | Versión publicada |
| Fecha de publicación: | 2005 |
| País: | Brasil |
| Institución: | Sociedade Brasileira de Patologia (SBP) |
| Repositorio: | Jornal Brasileiro de Patologia e Medicina Laboratorial (Online) |
| Idioma: | inglés |
| OAI Identifier: | oai:scielo:S1676-24442005000600006 |
| Acceso en línea: | http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1676-24442005000600006 |
| Access Level: | acceso abierto |
| Palabra clave: | Paraffin-embedded tissues DNA PCR |
| Sumario: | BACKGROUND: The most common human archival specimens are formalin-fixed, paraffin-embedded tissues (PETs). DNA can be extracted from PETs, but sometimes, it is unsuitable for molecular techniques as slow degradation of DNA occurs with time. OBJECTIVE: The aim of this study was to verify and discuss if samples of oral PETs archived for the past 40-years are possible substrates for molecular biology studies, using PCR. Methods: The samples were submitted to phenol-chloroform extraction method. DNA was qualified and quantified by spectrophotometer analysis, electrophoresis and amplification by PCR. RESULTS: It was observed a weak positive correlation between genomic DNA yield and specimen age. The agarose gel electrophoresis demonstrated that genomic DNA length was more frequently composed of small fragments. The 268-bp fragments of the beta-globin gene was amplified in 55% of cases and preferentially in more recent ones, which showed strong amplification if compared with older samples. WAF1 gene with 149-bp presented weak but detectable amplification in 75% of cases. The 536-bp fragment of beta-globin gene was detected in 25% of samples. The amplification was intense in genomic DNA extracted from recent cases and weak in older ones. CONCLUSION: This study shown that, despite degradation, it is possible to use genomic DNA obtained from PETs, archived for the past forty years, in PCR amplification of small DNA products, being large DNA fragments more difficult to amplificate. |
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