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

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17:38 min

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April 27, 2012

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

Chapters

  • 00:05Title
  • 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

Automatic Translation

Automatic Translation

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

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