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Radioaktivt In situ Hybridisering til påvisning af Forskellige genekspressionsmønstre i Tissue
JoVE Journal
Biology
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JoVE Journal Biology
Radioactive in situ Hybridization for Detecting Diverse Gene Expression Patterns in Tissue

Radioaktivt In situ Hybridisering til påvisning af Forskellige genekspressionsmønstre i 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

Denne protokol er med held anvendt til kvantitativ detektering niveauer og rumlige mønstre af mRNA-ekspression i forskellige vævstyper tværs vertebrate arter. Fremgangsmåden kan detektere lave tæthed transkripter og tillader behandling af hundredvis af objektglas samtidigt. Vi præsenterer denne protokol ved hjælp af udtryk profilering af aviær embryonale hjernens dannelse som et eksempel.

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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