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Studying Cell Cycle-regulated Gene Expression by Two Complementary Cell Synchronization Protocols
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JoVE 杂志 遗传学
Studying Cell Cycle-regulated Gene Expression by Two Complementary Cell Synchronization Protocols

Studying Cell Cycle-regulated Gene Expression by Two Complementary Cell Synchronization Protocols

DOI:
10.3791/55745-v

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12:02 min

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June 06, 2017

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, ,
1Department of Cell Biology and Histology,University of the Basque Country, UPV/EHU, 2Department of Genetics, Physical Anthropology and Animal Physiology,University of the Basque Country, UPV/EHU, 3Department of Molecular Mechanisms of Disease,University of Zurich

Chapters

  • 00:05Title
  • 00:41Thymidine- and Nocodazole-based (Thy-noc) Synchronization and Release
  • 03:34HU-based Synchronization and Release from G1/S Boundary
  • 05:14Collecting Samples for FACS Analysis and RNA Extraction
  • 07:00RNA Extraction for Gene Expression Analysis
  • 09:02Results: Thy-noc and HU Cell Synchronization Protocols Enable Analysis of Gene Expression During Specific Phases of the Cell Cycle
  • 11:07Conclusion

Summary

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We report two cell synchronization protocols that provide a context for studying events related to specific phases of the cell cycle. We show that this approach is useful for analyzing the regulation of specific genes in an unperturbed cell cycle or upon exposure to agents affecting the cell cycle.

Tags

Cell CycleGene ExpressionCell SynchronizationU2OS CellsThymidine BlockNocodazoleMitotic Cell ArrestG2-MRNA ExtractionFACS Analysis

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