Randomized terminal linker-dependent PCR: a versatile and sensitive method for detection of DNA damage / 生物医学与环境科学(英文)
Biomedical and Environmental Sciences
;
(12): 203-208, 2002.
Artículo
en Inglés
| WPRIM
| ID: wpr-264316
ABSTRACT
<p><b>OBJECTIVE</b>To design and develop a novel, sensitive and versatile method for in vivo foot printing and studies of DNA damage, such as DNA adducts and strand breaks.</p><p><b>METHODS</b>Starting with mammalian genomic DNA, single-stranded products were made by repeated primer extension, these products were ligated to a double-stranded linker having a randomized 3' overhang, and used for PCR. DNA breaks in p53 gene produced by restriction endonuclease AfaI were detected by using this new method followed by Southern hybridization with DIG-labeled probe.</p><p><b>RESULTS</b>This randomized terminal linker-dependent PCR (RDPCR) method could generate band signals many-fold stronger than conventional ligation-mediated PCR (LMPCR), and it was more rapid, convenient and accurate than the terminal transferase-dependent PCR (TDPCR).</p><p><b>CONCLUSION</b>DNA strand breakage can be detected sensitively in the gene level by RDPCR. Any lesion that blocks primer extension should be detectable.</p>
Texto completo:
Disponible
Índice:
WPRIM (Pacífico Occidental)
Asunto principal:
Daño del ADN
/
Reacción en Cadena de la Polimerasa
/
Genes p53
/
Sensibilidad y Especificidad
/
Cartilla de ADN
/
Aductos de ADN
/
Genética
/
Mamíferos
/
Métodos
Tipo de estudio:
Ensayo Clínico Controlado
/
Estudio diagnóstico
Límite:
Animales
/
Humanos
Idioma:
Inglés
Revista:
Biomedical and Environmental Sciences
Año:
2002
Tipo del documento:
Artículo
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