Rapid identification of eels Anguilla japonica and Anguilla anguilla by polymerase chain reaction with single nucleotide polymorphism-based specific probes
| Autor: | Gen Kaneko, Keijiro Sezaki, Misako Nakaya, Shugo Watabe, Hidehiro Kondo, Shiro Itoi |
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| Rok vydání: | 2005 |
| Předmět: |
biology
fungi food and beverages Single-nucleotide polymorphism Aquatic Science biology.organism_classification 16S ribosomal RNA Molecular biology Japonica law.invention Real-time polymerase chain reaction law TaqMan Restriction fragment length polymorphism Primer (molecular biology) Polymerase chain reaction |
| Zdroj: | Fisheries Science. 71:1356-1364 |
| ISSN: | 1444-2906 0919-9268 |
| DOI: | 10.1111/j.1444-2906.2005.01102.x |
| Popis: | Standard molecular techniques, such as sequencing and restriction fragment length polymorphism analysis after polymerase chain reaction (PCR) amplification are relatively complicated, and species identification can take a long time when using such techniques. We established a quick method, using PCR with species-specific TaqMan Minor Groove Binder (MGB) probes based on single nucleotide polymorphism (SNP) to distinguish the two eel species Anguilla japonica and Anguilla anguilla. This method can be used in processed products. Partial sequences of the mitochondrial 16S rRNA gene were compared between A. japonica and A. anguilla to design a primer pair common to both A. japonica and A. anguilla and probes specific to A. japonica and A. anguilla. Different fluorescence intensities were produced in two PCR microtubes each containing A. japonica and A. anguilla-specific probes for one target sample. We observed the fluorescence intensity of PCR products in microtubes under ultraviolet transillumination, with similar results to those obtained by real-time PCR. Therefore, SNP-based PCR is a powerful tool for identifying materials of processed foods from either A. japonica or A. anguilla. |
| Databáze: | OpenAIRE |
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