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Quick Fluorescent In Situ Hybridization Protocol for Xist RNA Combined with Immunofluorescence of Histone Modification in X-chromosome Inactivation
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Journal JoVE Biologie
Quick Fluorescent In Situ Hybridization Protocol for Xist RNA Combined with Immunofluorescence of Histone Modification in X-chromosome Inactivation

Quick Fluorescent In Situ Hybridization Protocol for Xist RNA Combined with Immunofluorescence of Histone Modification in X-chromosome Inactivation

DOI:
10.3791/52053-v

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12:42 min

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November 26, 2014

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, , , ,
1Division of Reproductive Sciences,Cincinnati Children’s Hospital Medical Center, 2Department of Pediatrics,University of Cincinnati College of Medicine

Chapitres

  • 00:05Titre
  • 02:21Preparing the FISH Oligonucleotide Probes
  • 04:05Preparing Slides for Immuno-FISH Using a Cytospin
  • 06:23Performing Immunoflourescence
  • 08:36Performing FISH
  • 10:34Results: Immuno–FISH Staining of Xist RNA and Histone H3K27me3
  • 11:55Conclusion

Summary

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Traduction automatique

We developed an easily customized strand-specific fluorescent in situ hybridization (FISH) protocol combined with immunofluorescence. This allows for a detailed examination of RNA dynamics with simultaneous insight into the chromatin structure, nuclear organization, and transcriptional regulation at the single cell level.

Tags

Keywords: Fluorescent In Situ HybridizationXist RNAImmunofluorescenceHistone ModificationX-chromosome InactivationLncRNAXist CloudH3K27me3Oligonucleotide ProbesImmuno-FISH

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