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Gecombineerde DNA-RNA-TL<em> In situ</em> Hybridisatie (FISH) aan X-chromosoom inactivatie studeren in Gedifferentieerde vrouwelijke muis embryonale stamcellen
JoVE Journal
Biology
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JoVE Journal Biology
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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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

Fluorescente in situ hybridisatie (FISH) maakt de detectie van nucleïnezuren in hun eigen omgeving binnen cellen. We beschrijven hier een protocol voor gecombineerde, gelijktijdige detectie van RNA en DNA door middel van FISH, die kan worden gebruikt om X-chromosoom inactivatie in muis embryonale stamcellen.

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