Optimization of fluorescence in situ hybridization (FISH) for the identification of two polar coccoid green algae species

高小艳; 李运广; 李会荣; 陈雯莉; 罗玮

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Optimization of fluorescence in situ hybridization (FISH) for the identification of two polar coccoid green algae species

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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) 10 mg·mL^(-1) lysozyme solution pretreatment at 37℃for 30 min;(2) the hybridization buffer including 20%formamide;(3) the hybridization condition was 47℃for 6 h.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.

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《极地科学进展：英文版》 |2010年第2期|167-179|共13页
作者
高小艳; 李运广; 李会荣; 陈雯莉; 罗玮;
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作者单位

College of Life Science and Technology;

State Key Laboratory of Agricultural Microbiology;

Huazhong Agricultural University;

Key Laboratory for Polar Science of the State Oceanic Administration;

Polar Research Institute of China;

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原文格式 PDF
正文语种 chi
中图分类植物细胞遗传学;氯霉素及其衍生物;
关键词
荧光原位杂交; 优化分离; 绿藻; 流式细胞术; 鱼; 鉴定; 种类; 球状;

机译：荧光原位杂交;优化分离;绿藻;流式细胞术;鱼;鉴定;种类;球状;

Optimization of fluorescence in situ hybridization (FISH) for the identification of two polar coccoid green algae species

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