Whole-Mount RNA In Situ Hybridization and Immunofluorescence of Xenopus Embryos and Tadpoles | Zendy

Helen Rankin Willsey | Zendy

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Whole-Mount RNA In Situ Hybridization and Immunofluorescence of Xenopus Embryos and Tadpoles

Author(s) -

Helen Rankin Willsey

Publication year - 2021

Publication title -

cold spring harbor protocols

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.674

H-Index - 51

eISSN - 1940-3402

pISSN - 1559-6095

DOI - 10.1101/pdb.prot105635

Subject(s) - xenopus , tadpole (physics) , in situ hybridization , immunofluorescence , embryo , rna , in situ , biology , microbiology and biotechnology , messenger rna , staining , computational biology , chemistry , gene , genetics , antibody , physics , organic chemistry , particle physics

A major advantage of experimentation in Xenopus is the ability to query the localization of endogenous proteins and RNAs in situ in the entire animal during all of development. Here I describe three variations of staining to visualize mRNAs and proteins in developing Xenopus embryos and tadpoles. The first section outlines a traditional colorimetric staining for mRNAs that is suitable for all stages of development, and the second extends this protocol for fluorescence-based detection for higher spatial and quantitative resolution. The final section details detection of proteins by immunofluorescence, optimized for tadpole stages but widely applicable to others. Finally, optimization strategies are provided.

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