PUBLICATION
Comparative in situ hybridization protocols in zebrafish
- Authors
- Dunn, K., Vashisht, A., Hammond-Weinberger, D.R.
- ID
- ZDB-PUB-220907-39
- Date
- 2022
- Source
- Biotechniques 73(3): 123-130 (Journal)
- Registered Authors
- Keywords
- Cabin1, atoh1b, comparative, gene expression analysis, protocol, stain, technique, zebrafish
- MeSH Terms
-
- Animals
- Chlorides
- Dextran Sulfate
- In Situ Hybridization
- Polyvinyl Alcohol*
- Zebrafish*/genetics
- PubMed
- 36065907 Full text @ Biotechniques
Citation
Dunn, K., Vashisht, A., Hammond-Weinberger, D.R. (2022) Comparative in situ hybridization protocols in zebrafish. Biotechniques. 73(3):123-130.
Abstract
In situ hybridization is a commonly used technique in molecular biology to assess the temporal and spatial expression of a given gene. As a long and labor-intensive protocol, double in situ hybridization, which detects two genes in series, is challenging and can require a lot of troubleshooting. Optional additives, polyvinyl alcohol and dextran sulfate, were tested in a standard in situ hybridization protocol and several colorimetric stain pairings using double in situ hybridization in zebrafish embryos. Optional additives can improve staining time and reduce nonspecific background. Nitro-blue tetrazolium chloride/5-bromo-4-chloro-3-indolyl phosphate (BCIP) + Fast Red/BCIP was the most effective stain pairing. As a proof-of-concept, this work shows that Cabin1 and atoh1b are expressed in distinct regions of the developing zebrafish brain.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping