Whole-mount in situ hibridization and detection of RNAs in vertebrate embryos and isolated organs chapter. “Whole-mount in situ hybridization with Xenopus embryos”

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UNIT 14.9     Whole-Mount In Situ Hybridization and Detection of RNAs in Vertebrate Embryos and Isolated Organs Anne Pizard (ISH in amniotic embryos and organs)1, Anna Haramis2, Andrés E. Carrasco (Xenopus embryos)3, Paula Franco (Xenopus embryos)3, Silvia López (Xenopus embryos)3, and Alejandra Paganelli (Xenopus embryos)3 1Harvard Medical School, Boston, Massachusetts 2EMBL, Heidelberg, Germany 3University of Buenos Aires-CONICET School of Medicine, Buenos Aires, Argentina Abstract Nonisotopic in situ hybridization using intact embryos or organs is an important method for determining the spatial distribution of RNAs. Because it allows the analysis of large numbers of samples, it is amenable to temporal expression studies and comparison between different genotypes. It offers sensitivity and reproducibility. In addition, histological details are not lost during the staining process. The protocols in this unit can be used for whole-mount in situ hybridization in Xenopus, mouse, and chicken embryos, as well as dissected organs from mouse and chicken. Preparation of digoxigenin-labeled riboprobes is also described. Keywords: in situ hybridization; gene expression; RNA; embryo Información suministrada por el agente en SIGEVA