Revelation of Effective Methods for Detection of Viral Nervous Necrosis Virus Gene using Polymerase Chain Reaction in Barfin Flounder, Verasper moseri.
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Accession number;01A0448118
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| Title;Revelation of Effective Methods for Detection of Viral Nervous Necrosis Virus Gene using Polymerase Chain Reaction in Barfin Flounder, Verasper moseri. |
| Author;
WATANABE KEN'ICHI
(Nihonsaibaigyogyokyo)
YOSHIMIZU MAMORU
(Hokkaido Univ., Graduate School of Fisheries Sci., JPN)
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Journal Title;Suisan Zoshoku
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Journal Code:Y0285A
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ISSN:0371-4217
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VOL.49;NO.1;PAGE.85-90(2001)
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| Figure&Table&Reference;TBL.2, REF.29 |
| Pub. Country;Japan |
| Language;Japanese |
| Abstract;Detection rate of viral nervous necrosis (VNN) virus gene using polymerase chain reaction was investigated. Tested specimens were: just hatched larvae, heads of larvae, eye or brain of juveniles, ovarian fluid and sperm obtained from brood fish. The specimens were mixed with a 10-fold serial dilution of virus solutions prepared from the eyes and brain of the Pacific Cod, Gadus macrocephalus, affected with VNN. For nucleic acid extraction, a comparison was made between 20-proteinase K, SDS- proteinase K, acid guanidium phenol chloroform, Isogen, TRIzol, RNA isolation kit, Catrimox-14, and High Pure Viral Nucleic Acid Kit. Isogen and/or RNA isolation kit showed the highest detection rate. PTC-200 and PJ 480 thermal cyclers were more effective than the PC-700 model. In comparison of reverse transcriptase, AMV, M-MLV, and Super Script II were tested; r Taq or Ex Taq was used as the DNA polymerase. Pairing of Super Script and Ex Taq was most effective. In PCR programs, 3-temperature PCR was more effective than 2-temperature PCR. (author abst.) |
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