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 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.
Adolph
S
S
Hameister
H
H
D000818:Animals; D001710:Biotin / Q000378:metabolism; D002460:Cell Line; D002871:Chromosome Banding; D004264:DNA Topoisomerases, Type I / Q000235:genetics*; D003850:Deoxyribonuclease I; D003856:Deoxyuracil Nucleotides / Q000378:metabolism; D005260:Female; D006801:Humans; D007621:Karyotyping; D008297:Male; D008677:Metaphase; D051379:Mice; D014176:Protein Biosynthesis; D011487:Protein Conformation; D014998:Y Chromosome
D003856:Deoxyuracil Nucleotides; C027078:deoxyuridine triphosphate; D001710:Biotin; D003850:Deoxyribonuclease I; D004264:DNA Topoisomerases, Type I