caa a22 4500 445319135 CHVBK 20180317142658.0 cr unu---uuuuu 170323e20110401xx s 000 0 eng 10.1007/s10126-010-9301-3 doi (NATIONALLICENCE)springer-10.1007/s10126-010-9301-3 Investigation on Natural Diets of Larval Marine Animals Using Peptide Nucleic Acid-Directed Polymerase Chain Reaction Clamping [Elektronische Daten] [Seinen Chow, Sayaka Suzuki, Tadashi Matsunaga, Shane Lavery, Andrew Jeffs, Haruko Takeyama] The stomach contents of the larvae of marine animals are usually very small in quantity and amorphous, especially in invertebrates, making morphological methods of identification very difficult. Nucleotide sequence analysis using polymerase chain reaction (PCR) is a likely approach, but the large quantity of larval (host) DNA present may mask subtle signals from the prey genome. We have adopted peptide nucleic acid (PNA)-directed PCR clamping to selectively inhibit amplification of host DNA for this purpose. The Japanese spiny lobster (Panulirus japonicus) and eel (Anguilla japonica) were used as model host and prey organisms, respectively. A lobster-specific PNA oligomer (20 bases) was designed to anneal to the sequence at the junction of the 18S rDNA gene and the internal transcribed spacer 1 (ITS1) of the lobster. PCR using eukaryote universal primers for amplifying the ITS1 region used in conjunction with the lobster-specific PNA on a mixed DNA template of lobster and eel demonstrated successful inhibition of lobster ITS1 amplification while allowing efficient amplification of eel ITS1. This method was then applied to wild-caught lobster larvae of P. japonicus and P. longipes bispinosus collected around Ryukyu Archipelago, Japan. ITS1 sequences of a wide variety of animals (Ctenophora, Cnidaria, Crustacea, Teleostei, Mollusca, and Chaetognatha) were detected. Springer Science+Business Media, LLC, 2010 PCR-clamping nationallicence PNA nationallicence Gut content analysis nationallicence Larval diet nationallicence Spiny lobster nationallicence Chow Seinen National Research Institute of Fisheries Science, 6-31-1 Nagai, 236-0316, Yokosuka, Japan aut Suzuki Sayaka Tokyo University of Agriculture and Technology, 2-24-16 Naka-cho, 184-8588, Koganei, Japan aut Matsunaga Tadashi Tokyo University of Agriculture and Technology, 2-24-16 Naka-cho, 184-8588, Koganei, Japan aut Lavery Shane Leight Marine Laboratory, University of Auckland, Box 349, Warkworth, 0941, Northland, New Zealand aut Jeffs Andrew Leight Marine Laboratory, University of Auckland, Box 349, Warkworth, 0941, Northland, New Zealand aut Takeyama Haruko Department of Life Science and Medical Bio-Science, Waseda University, 2-2 Wakamatsu-cho, 162-8480, Tokyo, Japan aut Marine Biotechnology Springer-Verlag 13/2(2011-04-01), 305-313 1436-2228 13:2<305 2011 13 10126 https://doi.org/10.1007/s10126-010-9301-3 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s10126-010-9301-3 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Chow Seinen National Research Institute of Fisheries Science, 6-31-1 Nagai, 236-0316, Yokosuka, Japan aut NATIONALLICENCE 700 1- Suzuki Sayaka Tokyo University of Agriculture and Technology, 2-24-16 Naka-cho, 184-8588, Koganei, Japan aut NATIONALLICENCE 700 1- Matsunaga Tadashi Tokyo University of Agriculture and Technology, 2-24-16 Naka-cho, 184-8588, Koganei, Japan aut NATIONALLICENCE 700 1- Lavery Shane Leight Marine Laboratory, University of Auckland, Box 349, Warkworth, 0941, Northland, New Zealand aut NATIONALLICENCE 700 1- Jeffs Andrew Leight Marine Laboratory, University of Auckland, Box 349, Warkworth, 0941, Northland, New Zealand aut NATIONALLICENCE 700 1- Takeyama Haruko Department of Life Science and Medical Bio-Science, Waseda University, 2-2 Wakamatsu-cho, 162-8480, Tokyo, Japan aut NATIONALLICENCE 773 0- Marine Biotechnology Springer-Verlag 13/2(2011-04-01), 305-313 1436-2228 13:2<305 2011 13 10126 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer