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.
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.
Mühlmann-Díaz
M C
MC
Colorado State University, Fort Collins 80523-1673, USA.
Dullea
R G
RG
Bedford
J S
JS
D000818:Animals; D017209:Apoptosis; D001973:Bromodeoxyuridine / Q000032:analysis; D016466:CHO Cells; D002875:Chromosomes / Q000737:chemistry / Q000648:ultrastructure*; D002880:Chromosomes, Human, Pair 11 / Q000737:chemistry; D006224:Cricetinae; D004247:DNA / Q000032:analysis*; D015342:DNA Probes; D006367:HeLa Cells; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D008214:Lymphocytes / Q000528:radiation effects / Q000648:ultrastructure; D008866:Microsurgery; D016133:Polymerase Chain Reaction
D015342:DNA Probes; D004247:DNA; D001973:Bromodeoxyuridine