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 | |
| 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.
Funding
This work was partially supported by NIH R01AR070313 to Ashley W. Seifert.
| Funders | Funder number |
|---|---|
| National Institutes of Health (NIH) | R01AR070313 |
| National Institutes of Health (NIH) |
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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