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A FACS-based Protocol to Isolate RNA from the Secondary Cells of Drosophila Male Accessory Glands
JoVE Journal
Genetica
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JoVE Journal Genetica
A FACS-based Protocol to Isolate RNA from the Secondary Cells of Drosophila Male Accessory Glands
DOI:

06:54 min

September 05, 2019

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Capitoli

  • 00:05Titolo
  • 01:03Accessory Gland Dissection
  • 02:50Tissue Digestion and Mechanical Cell Dissociation
  • 04:08Results: Representative Secondary Cell (SC) RNA Isolation and Analysis
  • 05:55Conclusion

Summary

Traduzione automatica

Here, we present a protocol to dissociate and sort a specific cell population from the Drosophila male accessory glands (secondary cells) for RNA sequencing and RT-qPCR. Cell isolation is accomplished through FACS purification of GFP-expressing secondary cells after a multistep-dissociation process requiring dissection, proteases digestion and mechanical dispersion.

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