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

15:54 min

June 14, 2014

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Capítulos

  • 00:05Título
  • 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

Tadução automática

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.

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