In Situ Hybridization Techniques for Paraffin-Embedded Adult Coral Samples
Corals are important ocean invertebrates that are critical for overall ocean health as well as human health. However, due to human impacts such as rising ocean temperatures and ocean acidification, corals are increasingly under threat. To tackle these challenges, advances in cell and molecular biolo...
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ftpubmed:oai:pubmedcentral.nih.gov:6235064 2023-05-15T17:51:52+02:00 In Situ Hybridization Techniques for Paraffin-Embedded Adult Coral Samples Traylor-Knowles, Nikki 2018-08-31 http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6235064/ http://www.ncbi.nlm.nih.gov/pubmed/30222153 https://doi.org/10.3791/57853 en eng MyJove Corporation http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6235064/ http://www.ncbi.nlm.nih.gov/pubmed/30222153 http://dx.doi.org/10.3791/57853 Copyright © 2018, Journal of Visualized Experiments J Vis Exp Retraction Text 2018 ftpubmed https://doi.org/10.3791/57853 2020-09-06T00:15:19Z Corals are important ocean invertebrates that are critical for overall ocean health as well as human health. However, due to human impacts such as rising ocean temperatures and ocean acidification, corals are increasingly under threat. To tackle these challenges, advances in cell and molecular biology have proven to be crucial for diagnosing the health of corals. Modifying some of the techniques commonly used in human medicine could greatly improve researchers' ability to treat and save corals. To address this, a protocol for in situ hybridization used primarily in human medicine and evolutionary developmental biology has been adapted for use in adult corals under stress. The purpose of this method is to visualize the spatial expression of an RNA probe in adult coral tissue that has been embedded in paraffin and sectioned onto glass slides. This method focuses on removal of the paraffin and rehydration of the sample, pretreatment of the sample to ensure permeability of the sample, pre-hybridization incubation, hybridization of the RNA probe, and visualization of the RNA probe. This is a powerful method when using non-model organisms to discover where specific genes are expressed, and the protocol can be easily adapted for other non-model organisms. However, the method is limited in that it is primarily qualitative, because expression intensity can vary depending on the amount of time used during the visualization step and the concentration of the probe. Furthermore, patience is required, as this protocol can take up to 5 days (and in many cases, longer) depending on the probe being used. Finally, non-specific background staining is common, but this limitation can be overcome. Text Ocean acidification PubMed Central (PMC) Patience ENVELOPE(-68.933,-68.933,-67.750,-67.750) Journal of Visualized Experiments 138 |
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Retraction Traylor-Knowles, Nikki In Situ Hybridization Techniques for Paraffin-Embedded Adult Coral Samples |
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Corals are important ocean invertebrates that are critical for overall ocean health as well as human health. However, due to human impacts such as rising ocean temperatures and ocean acidification, corals are increasingly under threat. To tackle these challenges, advances in cell and molecular biology have proven to be crucial for diagnosing the health of corals. Modifying some of the techniques commonly used in human medicine could greatly improve researchers' ability to treat and save corals. To address this, a protocol for in situ hybridization used primarily in human medicine and evolutionary developmental biology has been adapted for use in adult corals under stress. The purpose of this method is to visualize the spatial expression of an RNA probe in adult coral tissue that has been embedded in paraffin and sectioned onto glass slides. This method focuses on removal of the paraffin and rehydration of the sample, pretreatment of the sample to ensure permeability of the sample, pre-hybridization incubation, hybridization of the RNA probe, and visualization of the RNA probe. This is a powerful method when using non-model organisms to discover where specific genes are expressed, and the protocol can be easily adapted for other non-model organisms. However, the method is limited in that it is primarily qualitative, because expression intensity can vary depending on the amount of time used during the visualization step and the concentration of the probe. Furthermore, patience is required, as this protocol can take up to 5 days (and in many cases, longer) depending on the probe being used. Finally, non-specific background staining is common, but this limitation can be overcome. |
format |
Text |
author |
Traylor-Knowles, Nikki |
author_facet |
Traylor-Knowles, Nikki |
author_sort |
Traylor-Knowles, Nikki |
title |
In Situ Hybridization Techniques for Paraffin-Embedded Adult Coral Samples |
title_short |
In Situ Hybridization Techniques for Paraffin-Embedded Adult Coral Samples |
title_full |
In Situ Hybridization Techniques for Paraffin-Embedded Adult Coral Samples |
title_fullStr |
In Situ Hybridization Techniques for Paraffin-Embedded Adult Coral Samples |
title_full_unstemmed |
In Situ Hybridization Techniques for Paraffin-Embedded Adult Coral Samples |
title_sort |
in situ hybridization techniques for paraffin-embedded adult coral samples |
publisher |
MyJove Corporation |
publishDate |
2018 |
url |
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6235064/ http://www.ncbi.nlm.nih.gov/pubmed/30222153 https://doi.org/10.3791/57853 |
long_lat |
ENVELOPE(-68.933,-68.933,-67.750,-67.750) |
geographic |
Patience |
geographic_facet |
Patience |
genre |
Ocean acidification |
genre_facet |
Ocean acidification |
op_source |
J Vis Exp |
op_relation |
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6235064/ http://www.ncbi.nlm.nih.gov/pubmed/30222153 http://dx.doi.org/10.3791/57853 |
op_rights |
Copyright © 2018, Journal of Visualized Experiments |
op_doi |
https://doi.org/10.3791/57853 |
container_title |
Journal of Visualized Experiments |
container_issue |
138 |
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1766159152367796224 |