Analysis of DNA replication by fluorescence in situ hybridization

Methods : a Companion to Methods in Enzymology
B A Boggs, A C Chinault

Abstract

Fluorescence in situ hybridization (FISH) has been shown to discriminate between unreplicated and replicated regions of the genome in interphase nuclei, based on the number of specific fluorescent signals that can be detected. By examining the replication status of hybridizing sequences in large numbers of individual cells from an asynchronously growing population, it is possible to deduce a relative order of replication of different sequences. The availability of well-mapped genomic probes and the ability to compare results from different cell lines make this a convenient approach with which to map domains of replication timing control at any chromosomal position and to relate this to various patterns of gene expression. Since there appear to be important but poorly understood correlations among replication timing, chromatin structure, and transcriptional competence in mammalian cells, this provides a valuable approach to understanding these interrelationships at the molecular level. The procedures for using FISH to examine replication timing in mammalian nuclei are described here in detail, and the advantages and limitations of the approach are discussed. Some other strategies for using high-resolution FISH on chromatin fiber...Continue Reading

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Citations

Jun 30, 2010·Chromosoma·Elisa PalumboAntonella Russo
Sep 29, 1999·Proceedings of the National Academy of Sciences of the United States of America·R OfirS Selig
Jun 4, 2004·Genes & Development·Xiaohua WuDavid Livingston
Jun 8, 2002·Molecular and Cellular Biology·Jie ZhouCarl L Schildkraut
May 2, 2006·Molecular and Cellular Biology·Rebecca K Rowntree, Jeannie T Lee
Jun 9, 2004·International Journal of Cancer. Journal International Du Cancer·Zohar A DotanLydia Avivi
May 8, 2018·Genome Research·Juan Carlos Rivera-MuliaDavid M Gilbert
May 10, 2007·Nature Protocols·Véronique Azuara
Jul 10, 2018·Nucleic Acids Research·Dzmitry G BatrakouConrad A Nieduszynski
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Jan 2, 2003·Genetics in Medicine : Official Journal of the American College of Medical Genetics·Orit ReishAliza Amiel
Mar 12, 2020·Chromosome Research : an International Journal on the Molecular, Supramolecular and Evolutionary Aspects of Chromosome Biology·Beth A Sullivan
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Jan 27, 1998·Methods : a Companion to Methods in Enzymology·J R WuD M Gilbert
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