Whole-mount in situ hybridization using DIG-labeled probes in planarian

Methods Mol Biol. 2014:1211:41-51. doi: 10.1007/978-1-4939-1459-3_4.

Abstract

In recent years freshwater flatworms (planarian) have become a powerful model for studies of regeneration and stem cell biology. Whole-mount in situ hybridization (WISH) and fluorescent in situ hybridization (FISH) are key and most commonly used techniques to determine and visualize gene expression patterns in planaria. Here, we present the established version of whole-mount in situ hybridization (WISH) and whole-mount fluorescence in situ hybridization (WFISH) protocol optimized over the last years by several labs from the rapidly growing planaria field and give an overview of recently introduced modifications which can be critical in the study of low abundant transcripts.

MeSH terms

  • Animals
  • Base Sequence
  • Digoxigenin / analysis*
  • In Situ Hybridization, Fluorescence / methods*
  • Planarians / genetics*
  • RNA Probes / analysis*
  • RNA Probes / genetics
  • RNA, Helminth / analysis*
  • RNA, Helminth / genetics

Substances

  • RNA Probes
  • RNA, Helminth
  • Digoxigenin