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Blizard Institute - Faculty of Medicine and Dentistry
  1. Blizard Institute
  2. Research
  3. Core Facilities
  4. Flow Cytometry
  5. Uses of Flow Cytometry
  6. Cell Cycle Analysis
  7. Replication timing

Replication timing

Fractional S-phase sorting using bromodeoxyuridine uptake to study gene replication timing

The determination of the percentage of cells in S-phase is dependent upon the detection of a thymidine analogue, bromodeoxyuridine (BrdU), which when added to culture medium is incorporated into DNA during DNA replication is a well known technique in flow cytometry.

 Gene replication timing studies uses the BrdU labelling technique, S phase cells are sorted into 3 or more fractions. Collected fractions are analysed by PCR for genetic analysis after immunoprecipitation of DNA via incorporated BrdU.

  • S phase sorted fractions [new window]

Publications.

P Takousis, P Johonnett, J Williamson, P Sasieni, G Warnes, T Forshew, V Azuara, A Fisher, PJ Wu, PJ Jones, R Vatcheva, S Beck, D Sheer. Replication timing profile reflects the distinct functional and genomic features of the MHC Class II Region. Cell Cycle Oct 6:19, 2393-2398, 2007.

V Azuara, P Perry, S Sauer, M Spivakov, HF Jorgensen, RM John, M Gouti, M Casanova, G Warnes, M Merkenschlager, AG Fisher.Chromatin signatures of pluripotent cell lines. Nature Cell Biol. 8, 532-538, 2006.

P Perry, S Saur, N Billion, WD Richardson, M Spivakov, G Warnes, FJ Livesey, M Merkenschlager, AG Fisher, V Auara. A dynamic switch in the replication timing of key regulator genes in embryonic stem cells upon neural induction. Cell Cycle 3, (12), 1645-50, 2004.

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