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High Resolution Fluorescent In Situ Hybridization in Drosophila Embryos and Tissues Using Tyramide Signal Amplification
JoVE Journal
Genetics
This content is Free Access.
JoVE Journal Genetics
High Resolution Fluorescent In Situ Hybridization in Drosophila Embryos and Tissues Using Tyramide Signal Amplification
DOI:

14:21 min

October 19, 2017

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Chapters

  • 00:05Title
  • 01:09PCR Amplification of cDNA Templates from Plasmids
  • 03:43In vitro Transcription to Generate Antisense Probes
  • 05:25Drosophila Tissue Collection and Fixation
  • 07:51In Situ Hybridization
  • 09:05Probe Detection
  • 11:05Antibody Detection using Tyramide
  • 12:01Results: Robust Detection of RNA in Whole Drosophila Embryos and Dissected Tissues
  • 13:02Conclusion

Summary

Automatic Translation

The described RNA in situ hybridization protocol allows the detection of RNA in whole Drosophila embryos or dissected tissues. Using 96-well microtiter plates and tyramide signal amplification, transcripts can be detected at high resolution, sensitivity, and throughput, and at a relatively low cost.

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