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Application of 5-bromo-2'deoxyuridine as a label for in situ hybridization in chromosome microdissection and painting, and 3' OH DNA end labeling for apoptosis.
Mühlmann-Díaz, M C; Dullea, R G; Bedford, J S.
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  • Mühlmann-Díaz MC; Colorado State University, Fort Collins 80523-1673, USA.
Biotechniques ; 21(1): 82-6, 1996 Jul.
Article en En | MEDLINE | ID: mdl-8816240
We have utilized 5-bromo-2'deoxyuridine (BrdU) substituted DNA as a probe for a number of applications including, principally, for chromosome painting by fluorescence in situ hybridization (FISH) but also for DNA end-labeling to detect apoptotic cell death and for filter hybridization. Br-dUTP was used as a substitute for biotin or digoxigenin-dUTP in probe labeling techniques, such as random priming, nick translation, end-labeling or PCR. An especially useful application is that it may be incorporated into probe DNA while cells or plasmids in bacteria are growing in the presence of BrdU. This can be particularly advantageous when large quantities of probe are needed, since the cost per mole of digoxigenin-dUTP or biotin-dUTP is nearly 1000 times that of Br-dUTP. Also, if probe is prepared by growth in BrdU, the difference in cost to prepare equal quantities of labeled DNA is more than 10,000 times greater for biotin-dUTP.
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Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Bromodesoxiuridina / ADN / Cromosomas / Hibridación Fluorescente in Situ / Apoptosis Límite: Animals / Humans Idioma: En Revista: Biotechniques Año: 1996 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Reino Unido
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Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Bromodesoxiuridina / ADN / Cromosomas / Hibridación Fluorescente in Situ / Apoptosis Límite: Animals / Humans Idioma: En Revista: Biotechniques Año: 1996 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Reino Unido