Whole-mount in situ hybridization and detection of RNAs in vertebrate embryos and isolated organs.
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.
Pizard
Anne
A
Harvard Medical School, Boston, Massachusetts, USA.
Haramis
Anna
A
Carrasco
Andrés E
AE
Franco
Paula
P
López
Silvia
S
Paganelli
Alejandra
A
D000818:Animals; D002642:Chick Embryo; D004076:Digoxigenin / Q000032:analysis / Q000276:immunology; D004622:Embryo, Mammalian / Q000737:chemistry* / Q000648:ultrastructure; D004625:Embryo, Nonmammalian / Q000737:chemistry* / Q000648:ultrastructure; D007140:Immunoglobulin Fab Fragments / Q000032:analysis / Q000276:immunology; D017403:In Situ Hybridization / Q000379:methods*; D007202:Indicators and Reagents; D051379:Mice; D015347:RNA Probes; D012333:RNA, Messenger / Q000032:analysis*; D013045:Species Specificity; D013048:Specimen Handling / Q000379:methods; D014714:Vertebrates / Q000196:embryology; D014982:Xenopus laevis / Q000196:embryology
D007140:Immunoglobulin Fab Fragments; D007202:Indicators and Reagents; D015347:RNA Probes; D012333:RNA, Messenger; D004076:Digoxigenin