Whole-mount in situ hybridization detection of mRNA in GFP-marked drosophila imaginal disc mosaic clones

Adrienne VanZomeren-Dohm, Ellen Flannery, Molly Duman-Scheel

Research output: Contribution to journalArticle

6 Scopus citations

Abstract

Protocols for both generation of mosaic clones and whole-ount in situ hybridization detection of mRNA have led to an advanced understanding of Drosophila development. A simple means of combining these techniques for staining of imaginal discs would be useful, as it would allow for analysis of mRNA transcripts in marked mosaic clones. However, few researchers attempt such experiments due to the technical difficulty of simultaneously detecting clones and mRNA transcripts. Furthermore, maintaining the ability to use GFP-marked clones is desirable. However, typical Drosophila in situ hybridization protocols result in loss of GFP fluorescence. The method for double labeling of imaginal discs described here maintains the ability to identify mosaic clones, including those marked by GFP, while simultaneously allowing the detection of mRNA transcripts. The methodology described is technically manageable, robust, rapid, relatively inexpensive and of interest to many Drosophila researchers.

Original languageEnglish (US)
Pages (from-to)323-325
Number of pages3
JournalFly
Volume2
Issue number6
DOIs
StatePublished - Jan 1 2008

Keywords

  • Drosophila
  • GFP
  • Imaginal disc
  • In situ hybridization
  • Mosaic clone
  • MRNA detection

ASJC Scopus subject areas

  • Insect Science
  • Medicine(all)

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