DNA extraction from formalin-fixed laryngeal biopsies: Comparison of techniques.
Acta Otolaryngol
; 131(3): 330-3, 2011 Mar.
Article
em En
| MEDLINE
| ID: mdl-21142742
ABSTRACT
CONCLUSION:
PCR-quality DNA could be extracted from formalin-fixed paraffin-embedded (FFPE) samples with amplicons of at least 390 bp. Paraffin removal was not a necessary step. Proteinase K digestion was as efficient as the commercial kit for DNA extraction with a lower cost.OBJECTIVES:
To compare different DNA extraction protocols for FFPE samples and to describe the suitability of the extracted DNA for PCR reactions.METHODS:
For deparaffinization the following techniques were compared alkaline heat, xylene, and no removal. For DNA extraction, proteinase K digestion and organic extraction were compared. A commercial extraction kit was included as standard. DNA quality was assessed by PCR amplification of the HFE gene, for amplicons of 208 and 390 bp.RESULTS:
Extraction with the commercial kit and proteinase K digestion were more efficient than other techniques, with no statistical difference between them for both amplicons. The proteinase K digestion buffer had a cost of U$ 0.2 per sample and the commercial kit of U$7 per sample.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
DNA de Neoplasias
/
Neoplasias Laríngeas
/
Fixação de Tecidos
/
Inclusão em Parafina
Limite:
Humans
Idioma:
En
Revista:
Acta Otolaryngol
Ano de publicação:
2011
Tipo de documento:
Article