Mutational analysis using denaturing gradient gel electrophoresis and PCR.
Mutat Res
; 288(1): 103-12, 1993 Jul.
Article
en En
| MEDLINE
| ID: mdl-7686254
ABSTRACT
Denaturing gradient gel electrophoresis (DGGE) separates (DNA) molecules based on their sequence. Using the proper conditions, all base-pair substitutions can be resolved from the wild-type sequence using DGGE. Polymerase chain reaction (PCR) permits rapid amplification of a given region of the genome. In this paper, we demonstrate the utility of DGGE combined with PCR for mutation analysis by presenting different examples (i) analysis of mouse p53 cDNA for mutations, (ii) simultaneous analysis of thousands of 4NQO-induced mutants for mutations in HPRT exon 3, (iii) examination of the fidelity of the thermostable DNA polymerase isolated from Pyrococcus furiosus (Pfu), (iv) purification of mutant DNA from contaminating wild-type DNA from mouse spleenic T-cell clones.
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Colección:
01-internacional
Banco de datos:
MEDLINE
Asunto principal:
ADN de Cadena Simple
/
Análisis Mutacional de ADN
/
Reacción en Cadena de la Polimerasa
/
Electroforesis en Gel de Poliacrilamida
Límite:
Animals
/
Humans
Idioma:
En
Revista:
Mutat Res
Año:
1993
Tipo del documento:
Article