Whole-Mount In Situ Hybridization (WISH) for Salamander Embryos and Larvae

Sruthi Purushothaman; Ashley W. Seifert

doi:10.1007/978-1-0716-2659-7_5

Whole-Mount In Situ Hybridization (WISH) for Salamander Embryos and Larvae

Sruthi Purushothaman, Ashley W. Seifert

Research output: Chapter in Book/Report/Conference proceeding › Chapter › peer-review

1 Scopus citations

Abstract

Whole-mount in situ hybridization (WISH) is widely used to visualize transcribed gene sequences (mRNA) in developing embryos, larvae, and other nucleotide probe permeable tissue samples. This methodology involves the hybridization of an antisense nucleotide probe to the target mRNA, followed by chromogen or fluorescence-based detection. Here we describe a protocol for the spatiotemporal analysis of mRNA transcripts in axolotl embryos/larvae using digoxigenin-labeled riboprobes, anti-digoxigenin alkaline phosphatase, Fab fragments antibody, and NBT/BCIP chromogen detection.

Original language	English
Title of host publication	Methods in Molecular Biology
Pages	95-107
Number of pages	13
DOIs	https://doi.org/10.1007/978-1-0716-2659-7_5
State	Published - 2023

Publication series

Name	Methods in Molecular Biology
Volume	2562
ISSN (Print)	1064-3745
ISSN (Electronic)	1940-6029

Bibliographical note

Publisher Copyright:
© 2023, The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.

Keywords

Digoxigenin
In situ hybridization (ISH)
In vitro transcription
Limb development
Limb regeneration
Riboprobe
Salamander

ASJC Scopus subject areas

Molecular Biology
Genetics

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10.1007/978-1-0716-2659-7_5

Cite this

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KW - Limb development

KW - Limb regeneration

KW - Riboprobe

KW - Salamander

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Whole-Mount In Situ Hybridization (WISH) for Salamander Embryos and Larvae

Abstract

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Keywords

ASJC Scopus subject areas

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