Whole Mount In Situ Hybridization of Mid-Gestation Mouse Embryos

In situ hybridization is a powerful technique that allows the visualization of specific RNA species in biological samples in exquisite detail. It has been particularly well explored in the field of developmental genetics. The spatial and temporal patterns of RNA expression provide us with critical information on likely gene function during embryonic development, and often inform the decision on whether to attempt further gene manipulation approaches. Furthermore, once a mouse strain with altered gene function has been created, in situ hybridization is a critical tool for revealing how the development of embryos with the mutation differs from that of wild-type embryos, and thus infer the function of the altered gene. Here, a well-tested protocol used to visualize RNA expression in whole-mount mid-gestation mouse embryos ranging from 8.5 to 14.5 days post-coitum (dpc) is described. © 2020 Wiley Periodicals LLC. Basic Protocol 1: RNA probe synthesis Alternate Protocol: Preparation of DNA template by PCR Basic Protocol 2: Embryo dissection Basic Protocol 3: Whole mount in situ hybridization Support Protocol: Generation of embryo powder.