Abstract
This chapter describes a nonradioactive method for the localization of mRNA in whole mouse embryos. It employs riboprobes labeled with digoxigenin, a steroid-like moiety not found in animal tissue. Digoxigenin-containing probe is visualized with a conjugate of antidigoxigenin Fab and alkaline phosphatase and colorimetric staining. The results are visualized in three dimensions, hence subtle patterns can be visualized without laborious sectioning.
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References
Rosen, B. and Beddington, R. S. P. (1993) Whole-mount in situ hybridization in the mouse embryo: gene expression in three dimensions. TIG 9, 162–167.
Tautz, D. and Pfeile, C. (1989) A non-radioactive in situ hybridization method for the localization of specific RNA’s in Drosophila embryos reveals translational control of the segmentation gene hunchback. Chromosoma 98, 81–85.
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© 1994 Humana Press Inc., Totowa, NJ
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Rosen, B., Beddington, R. (1994). Detection of mRNA in Whole Mounts of Mouse Embryos Using Digoxigenin Riboprobes. In: Isaac, P.G. (eds) Protocols for Nucleic Acid Analysis by Nonradioactive Probes. Methods in Molecular Biology™, vol 28. Humana Press. https://doi.org/10.1385/0-89603-254-X:201
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DOI: https://doi.org/10.1385/0-89603-254-X:201
Publisher Name: Humana Press
Print ISBN: 978-0-89603-254-5
Online ISBN: 978-1-59259-515-0
eBook Packages: Springer Protocols