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Related Experiment Video

Updated: Mar 14, 2026

Chromosome Replicating Timing Combined with Fluorescent In situ Hybridization

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Published on: December 10, 2012

Analysis of Single-Locus Replication Timing in Asynchronous Cycling Cells.

Lo Sardo Federica¹

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

Methods in Molecular Biology (Clifton, N.J.)

|September 24, 2016

Summary

Genomic regions replicate at different times during S-phase, with early and late replicating regions alternating. This study details a method to analyze replication timing in Drosophila cells using BrdU labeling and cell sorting.

Keywords:

BrdU Early FACS sorting Late Mid Replication timing S-phase

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Genome-wide Determination of Mammalian Replication Timing by DNA Content Measurement

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Genome-wide Determination of Mammalian Replication Timing by DNA Content Measurement

Genome-wide Determination of Mammalian Replication Timing by DNA Content Measurement

Published on: January 19, 2017

Area of Science:

Molecular Biology
Genomics
Cell Biology

Background:

Eukaryotic genomes replicate asynchronously, with distinct early and late S-phase replication timing.
Genomic organization into structural domains influences replication timing, correlating with epigenetic states.
Understanding replication timing is crucial for comprehending genome stability and gene regulation.

Purpose of the Study:

To describe a robust protocol for analyzing specific locus replication timing in Drosophila.
To adapt existing methods for use in Drosophila embryonic cell lines (S2 and S3).

Main Methods:

Utilized bromodeoxyuridine (BrdU) labeling to mark newly synthesized DNA.
Employed Fluorescence-Activated Cell Sorting (FACS) to isolate distinct S-phase fractions (early, mid, late).
Applied the protocol to asynchronous cycling Drosophila embryonic cell lines.

Main Results:

Successfully established and validated a protocol for replication timing analysis in Drosophila S2 and S3 cells.
Demonstrated the feasibility of distinguishing early, mid, and late replicating genomic loci using BrdU and FACS.

Conclusions:

The described BrdU labeling and FACS sorting method provides a reliable approach for studying replication timing in Drosophila.
This protocol facilitates further investigation into the regulation and functional significance of replication timing in eukaryotic genomes.