Detection of small, single-copy genes on protein-G-banded chromosomes by electron microscopy
作者:
R. Fetni,
N. Lemieux,
B. Malfoy,
B. Dutrillaux,
P.-E. Messier,
C.-L. Richer,
期刊:
Cytogenetic and Genome Research
(Karger Available online 1992)
卷期:
Volume 60,
issue 3-4
页码: 187-189
ISSN:1424-8581
年代: 1992
DOI:10.1159/000133332
出版商: S. Karger AG
数据来源: Karger
摘要:
A method for the detection by electron microscopy of chromosome banding after in situ hybridization of small, nonradioactive DNA sequences is described. Typical high-resolution G-banding is produced by adding 5-bromodeoxyuridine (BrdU) during the last part of the S-phase and by applying a monoclonal antibody against the BrdU-substituted chromosome segments, followed by the addition of protein G, but no further treatment. A protocol for in situ hybridization of small, single-copy biotinylated DNA sequences and their detection by immunogold tagging on banded chromosomes is also described. This combined approach permits high-resolution mapping of small DNA sequences and should be useful in discriminating between neighboring DNA fragments.
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