Development of real-time PCR assay for genetic identification of the mottled skate, Beringraja pulchra
| Autor: | Hae Young Lee, Hyun-Su Jo, Ho Zoon Chae, Nam-Soo Cho, Min-Hee Kim, Yang-Han Lee, In Kwan Hwang, Dong-Ho Choi, Pil-Won Kang, Ki-Won Park |
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| Rok vydání: | 2015 |
| Předmět: |
Identification methods
Genetics Mitochondrial DNA biology Mottled skate Sequence Analysis DNA Real-Time Polymerase Chain Reaction biology.organism_classification DNA Mitochondrial Molecular biology Pathology and Forensic Medicine Electron Transport Complex IV Real-time polymerase chain reaction Species Specificity Specific primers GenBank TaqMan Animals Taq Polymerase Skates Fish DNA Probes Law DNA Primers Minor groove |
| Zdroj: | Forensic Science International. 255:80-84 |
| ISSN: | 0379-0738 |
| Popis: | The mottled skate, Beringraja pulchra is one of the commercially important fishes in the market today. However, B. pulchra identification methods have not been well developed. The current study reports a novel real-time PCR method based on TaqMan technology developed for the genetic identification of B. pulchra . The mitochondrial cytochrome oxidase subunit 1 (COI) nucleotide sequences of 29 B. pulchra , 157 skates and rays reported in GenBank DNA database were comparatively analyzed and the COI sequences specific to B. pulchra was identified. Based on this information, a system of specific primers and Minor Groove Binding (MGB) TaqMan probe were designed. The assay successfully discriminated in 29 specimens of B. pulchra and 27 commercial samples with unknown species identity. For B. pulchra DNA, an average Threshold Cycle (Ct) value of 19.1±0.1 was obtained. Among 27 commercial samples, two samples showed average Ct values 19.1±0.0 and 26.7±0.1, respectively and were confirmed to be B. pulchra based on sequencing. The other samples tested showed undetectable or extremely weak signals for the target fragment, which was also consistent with the sequencing results. These results reveal that the method developed is a rapid and efficient tool to identify B. pulchra and might prevent fraud or mislabeling during the distribution of B. pulchra products. |
| Databáze: | OpenAIRE |
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