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Analysis of Single-Locus Replication Timing in Asynchronous Cycling Cells

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Polycomb Group Proteins

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1480))

Abstract

In higher eucaryotes, not all the genome is replicated simultaneously: there are parts of the genome that replicate at the beginning of S-phase (early S-phase), others that are replicated later. In each cell, early replicating genomic regions are alternated to late-replicating regions. In general, eucaryotic genomes are organized into structural domains where genes showing the same epigenetic state replicate at the same time.

Here, we will describe the protocol that we routinely used for the analysis of replication timing of specific loci in Drosophila embryonic cell lines (S2 and S3) based on BrdU labeling and FACS sorting of different S-phase fractions (early, mid, late) of asynchronous cycling cells.

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Authors and Affiliations

  1. Oncogenomic and Epigenetic Unit, Regina Elena Cancer Institute, via Elio Chianesi 53, 00144, Rome, Italy

    Lo Sardo Federica

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  1. Lo Sardo Federica
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Correspondence to Lo Sardo Federica .

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Editors and Affiliations

  1. IBCN-CNR, Institute of Cell Biology and Neurobiology and Istituto Nazionale di Genetica Molecolare “Romeo ed Enrica Invernizzi”, Rome, Italy

    Chiara Lanzuolo

  2. Istituto Nazionale di Genetica Molecolare “Romeo ed Enrica Invernizzi”, Milan, Italy

    Beatrice Bodega

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Federica, L.S. (2016). Analysis of Single-Locus Replication Timing in Asynchronous Cycling Cells. In: Lanzuolo, C., Bodega, B. (eds) Polycomb Group Proteins. Methods in Molecular Biology, vol 1480. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6380-5_5

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  • DOI: https://doi.org/10.1007/978-1-4939-6380-5_5

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6378-2

  • Online ISBN: 978-1-4939-6380-5

  • eBook Packages: Springer Protocols

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