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Combined DNA-RNA Fluorescent In situ Hybridization (FISH) to Study X Chromosome Inactivation in Differentiated Female Mouse Embryonic Stem Cells
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Combined DNA-RNA Fluorescent In situ Hybridization (FISH) to Study X Chromosome Inactivation in Differentiated Female Mouse Embryonic Stem Cells

Combined DNA-RNA Fluorescent In situ Hybridization (FISH) to Study X Chromosome Inactivation in Differentiated Female Mouse Embryonic Stem Cells

DOI:
10.3791/51628-v

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15:54 min

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June 14, 2014

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1Department of Reproduction and Development,Erasmus MC – University Medical Center

Chapters

  • 00:05Title
  • 02:57Induction of Differentiation in Female Embryonic Stem Cells to Induce X Chromosome Inactivation
  • 04:55Fixation of Differentiated ES Cells for Subsequent DNA-RNA FISH Analysis
  • 05:33Labeling of DNA Probes for FISH Experiments
  • 08:02Permeabilization, Pre-treatment and Hybridization of Fixed Cells for Combined DNA-RNA FISH
  • 11:04Post-hybridization Washes and Antibody Mediated Detection of Probe
  • 13:21Results: X Chromosome Inactivation in Differentiating Female ES Cells is Visualized by Combined DNA-RNA FISH
  • 14:57Conclusion

Summary

Automatic Translation

Automatic Translation

Fluorescent in situ hybridization (FISH) allows the detection of nucleic acids in their native environment within cells. We here describe a protocol for the combined, simultaneous detection of RNA and DNA by means of FISH, which can be used to study X chromosome inactivation in mouse embryonic stem cells.

Tags

Keywords: DNA FISHRNA FISHCombined DNA-RNA FISHX Chromosome InactivationXist RNAFemale Mouse Embryonic Stem CellsGene Expression RegulationCytogeneticsCancer DiagnosticsGenome Aberrations

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