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

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

DOI:
10.3791/55157-v

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08:06 min

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January 19, 2017

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, , ,
1Dept. of Microbiology and Molecular Genetics, IMRIC, Faculty of Medicine,Hebrew University of Jerusalem, 2The Core Research Facility, IMRIC, Faculty of Medicine,Hebrew University of Jerusalem

Chapters

  • 00:05Title
  • 00:40Collection of Cells from Culture
  • 01:34Fixation
  • 02:31Propidium Iodide Staining
  • 03:39Sorting
  • 05:32DNA Sonication
  • 06:19Results: Measurements of S/G1 DNA Content Produce Detailed Time of Replication Maps
  • 07:34Conclusion

Summary

Automatic Translation

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We describe here a relatively fast and simple approach for mapping genome-wide mammalian replication timing, from cell isolation to the basic analysis of the sequencing results. A genomic map of a representative replication program will be provided following the protocol.

Tags

Keywords: Genome-wide Replication TimingDNA Content MeasurementReplication FieldReplication Timing ProgramAdherent CellsTrypsin-EDTACentrifugationEthanol FixationPropidium Iodide StainingFlow Cytometry561 Nm Laser

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