Comparative in situ hybridization protocols in zebrafish
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. Opt...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
| Published: |
Future Science Ltd
2022-09-01
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| Series: | BioTechniques |
| Subjects: | |
| Online Access: | https://www.future-science.com/doi/10.2144/btn-2022-0038 |
| _version_ | 1797995179350687744 |
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| author | Krista Dunn Apoorva Vashisht Dena R Hammond-Weinberger |
| author_facet | Krista Dunn Apoorva Vashisht Dena R Hammond-Weinberger |
| author_sort | Krista Dunn |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-04-11T09:56:20Z |
| format | Article |
| id | doaj.art-7b15749da85d4ae3b2f9f476c6f849b1 |
| institution | Directory Open Access Journal |
| issn | 0736-6205 1940-9818 |
| language | English |
| last_indexed | 2024-04-11T09:56:20Z |
| publishDate | 2022-09-01 |
| publisher | Future Science Ltd |
| record_format | Article |
| series | BioTechniques |
| spelling | doaj.art-7b15749da85d4ae3b2f9f476c6f849b12022-12-22T04:30:33ZengFuture Science LtdBioTechniques0736-62051940-98182022-09-0173312313010.2144/btn-2022-0038Comparative in situ hybridization protocols in zebrafishKrista Dunn0Apoorva Vashisht1Dena R Hammond-Weinberger21Department of Biology, Murray State University, Murray, KY, USA1Department of Biology, Murray State University, Murray, KY, USA1Department of Biology, Murray State University, Murray, KY, USAIn 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.https://www.future-science.com/doi/10.2144/btn-2022-0038atoh1bCabin1comparativegene expression analysisprotocolstain |
| spellingShingle | Krista Dunn Apoorva Vashisht Dena R Hammond-Weinberger Comparative in situ hybridization protocols in zebrafish BioTechniques atoh1b Cabin1 comparative gene expression analysis protocol stain |
| title | Comparative in situ hybridization protocols in zebrafish |
| title_full | Comparative in situ hybridization protocols in zebrafish |
| title_fullStr | Comparative in situ hybridization protocols in zebrafish |
| title_full_unstemmed | Comparative in situ hybridization protocols in zebrafish |
| title_short | Comparative in situ hybridization protocols in zebrafish |
| title_sort | comparative in situ hybridization protocols in zebrafish |
| topic | atoh1b Cabin1 comparative gene expression analysis protocol stain |
| url | https://www.future-science.com/doi/10.2144/btn-2022-0038 |
| work_keys_str_mv | AT kristadunn comparativeinsituhybridizationprotocolsinzebrafish AT apoorvavashisht comparativeinsituhybridizationprotocolsinzebrafish AT denarhammondweinberger comparativeinsituhybridizationprotocolsinzebrafish |