Extraction and processing of circulating DNA from large sample volumes using methylation on beads for the detection of rare epigenetic events.
Clin Chim Acta
; 425: 169-75, 2013 Oct 21.
Article
em En
| MEDLINE
| ID: mdl-23911908
ABSTRACT
The use of methylated tumor-specific circulating DNA has shown great promise as a potential cancer biomarker. Nonetheless, the relative scarcity of tumor-specific circulating DNA presents a challenge for traditional DNA extraction and processing techniques. Here we demonstrate a single tube extraction and processing technique dubbed "methylation on beads" that allows for DNA extraction and bisulfite conversion for up to 2 ml of plasma or serum. In comparison to traditional techniques including phenol chloroform and alcohol extraction, methylation on beads yields a 1.5- to 5-fold improvement in extraction efficiency. The technique results in far less carryover of PCR inhibitors yielding analytical sensitivity improvements of over 25-fold. The combination of improved recovery and sensitivity make possible the detection of rare epigenetic events and the development of high sensitivity epigenetic diagnostic assays.
Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
DNA
/
Biomarcadores Tumorais
/
Reação em Cadeia da Polimerase
/
Carcinoma Pulmonar de Células não Pequenas
/
Proteínas Supressoras de Tumor
/
Neoplasias Pulmonares
Tipo de estudo:
Diagnostic_studies
Limite:
Aged
/
Female
/
Humans
/
Male
/
Middle aged
Idioma:
En
Revista:
Clin Chim Acta
Ano de publicação:
2013
Tipo de documento:
Article