In situ nick translation of metaphase chromosomes with biotin-labeled d-UTP

Limited nick translation experiments on fixed chromosomes were performed. Sites of preferential DNase-I nicking were made visible by the incorporation of biotin-labeled dUTP and subsequent binding of the streptavidin-peroxidase complex. This procedure leads to a banding pattern on the chromosomes wh...

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Bibliographic Details
Published in:Human genetics 1985-02, Vol.69 (2), p.117-121
Main Authors: Adolph, S, Hameister, H
Format: Article
Language:English
Subjects:
Animals
Biotin - metabolism
Cell Line
Chromosome Banding
Deoxyribonuclease I
Deoxyuracil Nucleotides - metabolism
DNA Topoisomerases, Type I - genetics
Female
Humans
Karyotyping
Male
Metaphase
Mice
Protein Biosynthesis
Protein Conformation
Y Chromosome
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Summary:Limited nick translation experiments on fixed chromosomes were performed. Sites of preferential DNase-I nicking were made visible by the incorporation of biotin-labeled dUTP and subsequent binding of the streptavidin-peroxidase complex. This procedure leads to a banding pattern on the chromosomes which is strongly DNase-I concentration dependent. Along the chromosome arms, regions of enhanced DNase-I sensitivity alternate with regions of lower DNase-I sensitivity. No complete G- or R-type banding pattern was observed. The easily identifiable human Y chromosome was studied more intensively. Compiled data show the heterochromatin of the Y chromosome stained as heavily as the euchromatin. The boundary between the eu- and heterochromatin on the long arm appears to be a site of preferential DNase-I sensitivity.
ISSN:0340-6717
1432-1203
DOI:10.1007/BF00293280