Unit

UNIT 14.9 Whole-Mount In Situ Hybridization and Detection of RNAs in Vertebrate Embryos and Isolated Organs

  1. Anne Pizard (ISH in amniotic embryos and organs)1,
  2. Anna Haramis2,
  3. Andrés E. Carrasco (Xenopus embryos)3,
  4. Paula Franco (Xenopus embryos)3,
  5. Silvia López (Xenopus embryos)3,
  6. Alejandra Paganelli (Xenopus embryos)3

Published Online: 1 MAY 2004

DOI: 10.1002/0471142727.mb1409s66

Current Protocols in Molecular Biology

Current Protocols in Molecular Biology

How to Cite

Pizard, A., Haramis, A., Carrasco, A. E., Franco, P., López, S. and Paganelli, A. 2004. Whole-Mount In Situ Hybridization and Detection of RNAs in Vertebrate Embryos and Isolated Organs. Current Protocols in Molecular Biology. 66:14.9:14.9.1–14.9.24.

Author Information

  1. 1

    Harvard Medical School, Boston, Massachusetts

  2. 2

    EMBL, Heidelberg, Germany

  3. 3

    University of Buenos Aires-CONICET School of Medicine, Buenos Aires, Argentina

Publication History

  1. Published Online: 1 MAY 2004
  2. Published Print: APR 2004

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