Optimization of fluorescence in situ hybridization (FISH) for the identification of two polar coccoid green algae species
Standard FISH protocols using fluorochrome-labeled oligonucleotide probes have been successfully applied for in situ detection. However, optimized protocols of FISH for specific eukaryotes in marine environments are often not developed. This study optimized the conditions of fluorescence in situ hyb...
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Polar Research Institute of China - PRIC
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ftarcticportal:oai:generic.eprints.org:2402 2023-10-09T21:44:11+02:00 Optimization of fluorescence in situ hybridization (FISH) for the identification of two polar coccoid green algae species Xiaoyan, Gao Yunguang, Li Huirong, Li Wenli, Chen Wei, Luo 2010-12 application/pdf http://library.arcticportal.org/2402/ http://library.arcticportal.org/2402/1/A2010-016.pdf en eng Polar Research Institute of China - PRIC http://library.arcticportal.org/2402/1/A2010-016.pdf Xiaoyan, Gao and Yunguang, Li and Huirong, Li and Wenli, Chen and Wei, Luo (2010) Optimization of fluorescence in situ hybridization (FISH) for the identification of two polar coccoid green algae species. Advances in Polar Science, 21 (2). pp. 167-179. Fauna Oceans Article PeerReviewed 2010 ftarcticportal 2023-09-13T22:54:14Z Standard FISH protocols using fluorochrome-labeled oligonucleotide probes have been successfully applied for in situ detection. However, optimized protocols of FISH for specific eukaryotes in marine environments are often not developed. This study optimized the conditions of fluorescence in situ hybridization(FISH) by using two polar isolated microalgae. The modified conditions were as follows: (1)10mg·mL-1 lysozyme solution pretreatment at 37°C for 30 min; (2)the hybridization buffer including 20% formamide; (3)the hybridization condition was 47°C for 6h. The cells enumerated by FISH were compared with those enumerated by flow cytometry(FCM) and DAPI to confirm the cell loss and hybridization efficiency. The optimized protocol was also successfully applied to Arctic Ocean samples, which were found to be dominated by Micromonas sp. The modified protocol showed a high relative efficiency and could be successfully applied for the detection of specific microbial eukaryotes in environmental samples. Article in Journal/Newspaper Advances in Polar Science Arctic Arctic Ocean Polar Science Polar Science Arctic Portal Library Arctic Arctic Ocean |
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Arctic Portal Library |
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ftarcticportal |
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English |
topic |
Fauna Oceans |
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Fauna Oceans Xiaoyan, Gao Yunguang, Li Huirong, Li Wenli, Chen Wei, Luo Optimization of fluorescence in situ hybridization (FISH) for the identification of two polar coccoid green algae species |
topic_facet |
Fauna Oceans |
description |
Standard FISH protocols using fluorochrome-labeled oligonucleotide probes have been successfully applied for in situ detection. However, optimized protocols of FISH for specific eukaryotes in marine environments are often not developed. This study optimized the conditions of fluorescence in situ hybridization(FISH) by using two polar isolated microalgae. The modified conditions were as follows: (1)10mg·mL-1 lysozyme solution pretreatment at 37°C for 30 min; (2)the hybridization buffer including 20% formamide; (3)the hybridization condition was 47°C for 6h. The cells enumerated by FISH were compared with those enumerated by flow cytometry(FCM) and DAPI to confirm the cell loss and hybridization efficiency. The optimized protocol was also successfully applied to Arctic Ocean samples, which were found to be dominated by Micromonas sp. The modified protocol showed a high relative efficiency and could be successfully applied for the detection of specific microbial eukaryotes in environmental samples. |
format |
Article in Journal/Newspaper |
author |
Xiaoyan, Gao Yunguang, Li Huirong, Li Wenli, Chen Wei, Luo |
author_facet |
Xiaoyan, Gao Yunguang, Li Huirong, Li Wenli, Chen Wei, Luo |
author_sort |
Xiaoyan, Gao |
title |
Optimization of fluorescence in situ hybridization (FISH) for the identification of two polar coccoid green algae species |
title_short |
Optimization of fluorescence in situ hybridization (FISH) for the identification of two polar coccoid green algae species |
title_full |
Optimization of fluorescence in situ hybridization (FISH) for the identification of two polar coccoid green algae species |
title_fullStr |
Optimization of fluorescence in situ hybridization (FISH) for the identification of two polar coccoid green algae species |
title_full_unstemmed |
Optimization of fluorescence in situ hybridization (FISH) for the identification of two polar coccoid green algae species |
title_sort |
optimization of fluorescence in situ hybridization (fish) for the identification of two polar coccoid green algae species |
publisher |
Polar Research Institute of China - PRIC |
publishDate |
2010 |
url |
http://library.arcticportal.org/2402/ http://library.arcticportal.org/2402/1/A2010-016.pdf |
geographic |
Arctic Arctic Ocean |
geographic_facet |
Arctic Arctic Ocean |
genre |
Advances in Polar Science Arctic Arctic Ocean Polar Science Polar Science |
genre_facet |
Advances in Polar Science Arctic Arctic Ocean Polar Science Polar Science |
op_relation |
http://library.arcticportal.org/2402/1/A2010-016.pdf Xiaoyan, Gao and Yunguang, Li and Huirong, Li and Wenli, Chen and Wei, Luo (2010) Optimization of fluorescence in situ hybridization (FISH) for the identification of two polar coccoid green algae species. Advances in Polar Science, 21 (2). pp. 167-179. |
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1779319941562892288 |