Whole-Mount In Situ Hybridization (WISH) Optimized for Gene Expression Analysis in Mouse Embryos and Embryoid Bodies
Whole-mount in situ hybridization (WISH) is a technique widely used in developmental biology to study the localization of RNA sequences in intact tissues or whole organisms. In this chapter we present a detailed protocol that was optimized for gene expression analysis in early stage mouse embryos (5.5–10.5 days post-coitum) and embryoid bodies formed by differentiating embryonic stem cells and can be used for the detection of up to two distinct RNA sequences simultaneously. The initial steps of the procedure are the generation of the labeled riboprobe(s) and the embryo or embryoid body preparation, which can be completed in less than 2 days. The actual WISH procedure, comprised of the hybridization, the post-hybridization washes, and the immunological staining, can be completed in 3 days.
Key wordsWISH In situ hybridization mRNA Mouse embryo Embryoid bodies Gene expression pattern
Eleni Dakou is funded by the Institute for the Promotion of Innovation through Science and Technology in Flanders (IWT-Vlaanderen). This research is supported by an Interuniversity Attraction Pole grant (IAP-P7-07).
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