TY - JOUR
T1 - Species identification of Anguilla japonica by real-time PCR based on a sequence detection system
T2 - A practical application to eggs and larvae
AU - Minegishi, Yuki
AU - Yoshinaga, Tatsuki
AU - Aoyama, Jun
AU - Tsukamoto, Katsumi
N1 - Funding Information:
We thank Yoshiaki Yamada of Irago Institute for providing the eggs. The study was partly supported by Grants-in-Aid for Creative Scientific Research No. 12NP0201 (DOBIS) from the Ministry of Education, Culture, Sports, Science and Technology of Japan and the Eel Research Foundation from “Nobori-kai”.
PY - 2009/10
Y1 - 2009/10
N2 - To develop a practical method for identifying Japanese eel Anguilla japonica eggs and larvae to species by a sequence detection system using a real-time polymerase chain reaction (PCR), we examined (i) the sensitivity of the system using samples at various developmental stages, and (ii) influences of intra- and interspecific DNA sequence variations in the PCR target region. PCR amplifications with extracted DNA solution at 7.0 ng l-1 or lower were efficient at distinguishing A. japonica from other anguillids. A single egg at the gastrula or later developmental stages could also be identified. Two sequence variations in the PCR target region were observed in 2 out of 35 A. japonica collected from three localities, and from four year classes at a single locality. These mutations, however, did not affect the result of species identification achieved by A. japonica-specific PCR primers and probe. The accuracy of this PCR-based method of species identification will help in field surveys of the species.
AB - To develop a practical method for identifying Japanese eel Anguilla japonica eggs and larvae to species by a sequence detection system using a real-time polymerase chain reaction (PCR), we examined (i) the sensitivity of the system using samples at various developmental stages, and (ii) influences of intra- and interspecific DNA sequence variations in the PCR target region. PCR amplifications with extracted DNA solution at 7.0 ng l-1 or lower were efficient at distinguishing A. japonica from other anguillids. A single egg at the gastrula or later developmental stages could also be identified. Two sequence variations in the PCR target region were observed in 2 out of 35 A. japonica collected from three localities, and from four year classes at a single locality. These mutations, however, did not affect the result of species identification achieved by A. japonica-specific PCR primers and probe. The accuracy of this PCR-based method of species identification will help in field surveys of the species.
KW - Anguilla japonica
KW - Cycles at threshold (Ct)
KW - Sequence detection system
KW - Species identification
KW - Template DNA concentrations
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U2 - 10.1093/icesjms/fsp158
DO - 10.1093/icesjms/fsp158
M3 - Article
AN - SCOPUS:70149121605
VL - 66
SP - 1915
EP - 1918
JO - ICES Journal of Marine Science
JF - ICES Journal of Marine Science
SN - 1054-3139
IS - 9
ER -