Comparative in situ hybridization protocols in zebrafish
| Autor: | Krista Dunn, Apoorva Vashisht, Dena R Hammond-Weinberger |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2022 |
| Předmět: | |
| Zdroj: | BioTechniques, Vol 73, Iss 3, Pp 123-130 (2022) |
| Druh dokumentu: | article |
| ISSN: | 1940-9818 0736-6205 |
| DOI: | 10.2144/btn-2022-0038 |
| Popis: | 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. |
| Databáze: | Directory of Open Access Journals |
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