A new and rapid method for visualising DNA replication in spread DNA by immunofluorescence detection of incorporated 5-iododeoxyuridine.
We have developed a new and rapid immunofluorescent method for visualisation of replicated regions on fixed DNA fibers. Using this method we have found in 5-fluorodeoxyuridine (FrdU)-blocked human cells distinct replication units covering about 60 kb of DNA and corresponding in size to single replicons or chromatin loops. Our results also suggest that nonadjacent replicons within a replicon cluster may be activated after FrdU arrest and that the method may be adapted for localization on fibers of specific DNA sequences.
Tomilin
N
N
Institute of Cytology of the Russian Academy of Sciences, St. Petersburg.
Rosanov
Y u
Yu
Zenin
V
V
Bozhkov
V
V
Vig
B
B
D002877:Chromosomes, Human / Q000648:ultrastructure; D004247:DNA / Q000032:analysis* / Q000096:biosynthesis; D004261:DNA Replication / Q000187:drug effects; D004273:DNA, Neoplasm / Q000032:analysis* / Q000096:biosynthesis; D005467:Floxuridine / Q000494:pharmacology; D005455:Fluorescent Antibody Technique; D006801:Humans; D007065:Idoxuridine; D017403:In Situ Hybridization / Q000379:methods; D007202:Indicators and Reagents; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive; D009691:Nucleic Acid Denaturation; D014407:Tumor Cells, Cultured
D004273:DNA, Neoplasm; D007202:Indicators and Reagents; D005467:Floxuridine; D004247:DNA; D007065:Idoxuridine