Journal
/
Biologie
/
Radioactive in situ Hybridization for Detecting Diverse Gene Expression Patterns in Tissue
Journal JoVE
Biologie
This content is Free Access.
Journal JoVE Biologie
Radioactive in situ Hybridization for Detecting Diverse Gene Expression Patterns in Tissue

Radioactive in situ Hybridization for Detecting Diverse Gene Expression Patterns in Tissue

DOI:
10.3791/3764-v

•

17:38 min

•

April 27, 2012

•

, ,
1Howard Hughes Medical Institute, Department of Neurobiology,Duke University , 2Department of Biological Sciences,Hokkaido University

Chapitres

  • 00:05Titre
  • 02:16Tissue Preparation
  • 03:20Pre-hybridization Treatment of Tissue
  • 04:36Probe Hybridization
  • 07:34Visualization of the Radioactive Signal
  • 10:59Generating Darkfield Color Images
  • 11:38Results: mRNA Expression Patterns in Avian Brain
  • 16:56Conclusion

Summary

Traduction automatique

Traduction automatique

This protocol is successfully used to quantitatively detect levels and spatial patterns of mRNA expression in multiple tissue types across vertebrate species. The method can detect low abundance transcripts and allows processing of hundreds of slides simultaneously. We present this protocol using expression profiling of avian embryonic brain formation as an example.

Tags

Radioactive In Situ HybridizationGene Expression PatternsTimingLevelCellular LocalizationCell TypeMRNAComplementary RNA ProbesRadioactive NucleotidesDetection SensitivityQuantitative AnalysisLight Sensitive X-ray FilmEmulsion CoatedTarget Gene ExpressionNon-radioactive Probe MethodsDIG-labelingAmplification StepsHRP-antibodiesTSA KitSignal IntensityTargeted MRNA AmountsQuantitative AnalysisDevelopmental Stages Of Embryos

Article

Embed

AJOUTER À LA PLAYLIST

Usage Statistics

Vidéos Connexes

The Preparation of Primary Hematopoietic Cell Cultures From Murine Bone Marrow for Electroporation

Non-radioactive in situ Hybridization Protocol Applicable for Norway Spruce and a Range of Plant Species

Using an Automated Cell Counter to Simplify Gene Expression Studies: siRNA Knockdown of IL-4 Dependent Gene Expression in Namalwa Cells

In vitro Reconstitution of the Active T. castaneum Telomerase

A High Throughput in situ Hybridization Method to Characterize mRNA Expression Patterns in the Fetal Mouse Lower Urogenital Tract

Analysis of Single-cell Gene Transcription by RNA Fluorescent In Situ Hybridization (FISH)

Visualization and Analysis of mRNA Molecules Using Fluorescence In Situ Hybridization in Saccharomyces cerevisiae

High Resolution Whole Mount In Situ Hybridization within Zebrafish Embryos to Study Gene Expression and Function

MicroRNA In situ Hybridization for Formalin Fixed Kidney Tissues

Measurement of Metabolic Rate in Drosophila using Respirometry

Read Article