PUBLICATION
Genotyping and Quantification of In Situ Hybridization Staining in Zebrafish
- Authors
- Dobrzycki, T., Krecsmarik, M., Monteiro, R.
- ID
- ZDB-PUB-200221-20
- Date
- 2020
- Source
- Journal of visualized experiments : JoVE (155): (Journal)
- Registered Authors
- Monteiro, Rui
- Keywords
- none
- MeSH Terms
-
- Animals
- Genotype
- In Situ Hybridization/methods*
- Zebrafish
- PubMed
- 32065138 Full text @ J. Vis. Exp.
Citation
Dobrzycki, T., Krecsmarik, M., Monteiro, R. (2020) Genotyping and Quantification of In Situ Hybridization Staining in Zebrafish. Journal of visualized experiments : JoVE. (155):.
Abstract
In situ hybridization (ISH) is an important technique that enables researchers to study mRNA distribution in situ and has been a critical technique in developmental biology for decades. Traditionally, most gene expression studies relied on visual evaluation of the ISH signal, a method that is prone to bias, particularly in cases where sample identities are known a priori. We have previously reported on a method to circumvent this bias and provide a more accurate quantification of ISH signals. Here, we present a simple guide to apply this method to quantify the expression levels of genes of interest in ISH-stained embryos and correlate that with their corresponding genotypes. The method is particularly useful to quantify spatially restricted gene expression signals in samples of mixed genotypes and it provides an unbiased and accurate alternative to the traditional visual scoring methods.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping