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Kombinerad DNA-RNA Fluorescent<em> In situ</em> Hybridisering (FISH) för att studera X-kromosom inaktivering i Differentierade Kvinna Mouse embryonala stamceller
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

Fluorescent in situ-hybridisering (FISH) tillåter detektion av nukleinsyror i deras nativa omgivning inom celler. Vi beskriver här ett protokoll för det kombinerade, samtidig upptäckt av RNA och DNA med hjälp av FISH, som kan användas för att studera X-kromosom inaktivering i mus embryonala stamceller.

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