A quantitative DNA methylation assay using mismatch hybridization and chemiluminescence / 生物医学与环境科学(英文)
Biomedical and Environmental Sciences
;
(12): 48-52, 2005.
Artículo
en Inglés
| WPRIM
| ID: wpr-329602
ABSTRACT
<p><b>OBJECTIVE</b>To develop a quantitative method for methylation analysis of the p16 gene based on mismatch hybridization and chemiluminescence.</p><p><b>METHODS</b>Genomic DNA was modified by sodium bisulfite to convert all unmethylated but not methylated cytosines to uracil, and subsequently a pair of primer having no CpG sites was designed for amplification target DNA containing methylated or unmethylated CpG sites. The PCR product spanning CpG sites were hybridized with two oligonucleotide probes which perfectly matched the methylated and unmethylated CpG sequences respectively, and the hybrids were detected by chemiluminescent method. The percentage of methylated target sequences could be estimated by calculating the ratio of signals obtained with two probes.</p><p><b>RESULTS</b>The percentage of methylation of artificial mixtures DNA showed a linear relation. There was a negative correlation between the methyaltion index with p16 transcriptional mRNA of p16 gene in tumor cell lines.</p><p><b>CONCLUSION</b>Compared with existing methods, this assay is nonisotopic, rapid, simple, and can be widely applied to the study of DNA methylation.</p>
Texto completo:
Disponible
Índice:
WPRIM (Pacífico Occidental)
Asunto principal:
Sulfitos
/
Regiones Promotoras Genéticas
/
Islas de CpG
/
Metilación de ADN
/
Genes p16
/
Reacción en Cadena de la Polimerasa de Transcriptasa Inversa
/
Línea Celular Tumoral
/
Mediciones Luminiscentes
/
Hibridación de Ácido Nucleico
Límite:
Humanos
Idioma:
Inglés
Revista:
Biomedical and Environmental Sciences
Año:
2005
Tipo del documento:
Artículo
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