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Primers to block the amplification of symbiotic apostome ciliate 18S rRNA gene in a PCR-based copepod diet study |
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| Authors: | Xiaoyan Yi Huan Zhang Guangxing Liu |
| Institution: | 1. College of Environmental Science and Engineering, Ocean University of China, Qingdao, 266100, China 2. Department of Marine Sciences, Connecticut University, Groton, Connecticut, 06340, USA 3. Ministry of Education, Key Laboratory of Marine Environment and Ecology (Ocean University of China), Qingdao, 266100, China |
| Abstract: | Pelagic copepods play an important role in the marine food web. However, a full understanding of the ecological status of this zooplankton group depends on the careful study of their natural diets. In previous PCR-based copepod diet studies, we found many apostome ciliates that live symbiotically under the exoskeleton of the copepods, and their sequences were often over-represented in the 18S rRNA gene (18S rDNA) libraries. As a first step to address this issue, we designed three apostome ciliate 18S rDNA blocking primers, and tested their blocking efficiency against apostome ciliate 18s rDNA under various PCR conditions. Using a semi-quantitative PCR method, we optimized the conditions to efficiently amplify the 18S rDNA of the prey while simultaneously excluding the symbiotic apostome ciliates. This technique will facilitate PCR-based diet studies of copepods and other zooplankton in their natural environments. |
| Keywords: | copepod 18S rDNA apostome ciliate blocking primer PCR |
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