연어와 무지개송어 판별을 위한 Ultra-fast PCR 판별법 개발 및 검증

다국어 초록 (Multilingual Abstract)

In this study, Ultra-fast polymerase chain (Ultra-fast PCR) assay was developed and verified to distinguish Oncorhynchus keta and Oncorhynchus mykiss which are difficult to be distinguished morphologically. Species-specific primers were designed for the CO I genes of mtDNA. The positive Ct value of O. keta was less than 24 cycles and the positive Ct value of O. mykiss was less than 20 cycles. The species-specific primers designed in this study for O. keta and O. mykiss were selectively amplified by O. keta and O. mykiss DNA, respectively. The sensitivity of O. keta and O. mykiss primer was 1 ng/μL. As a result of quantitative testing, it was confirmed that the “Guidelines on standard procedures for preparing analysis method such as food.” proposed by the Korea food and drug administration were met. A total of 14 monitoring samples, including 8 O. keta and 6 O. mykiss, showed no forgery and alteration cases. Ultra-fast PCR method developed and verified in this study for discrimination of O. keta and O. mykiss has considerable advantages in terms of speed and field applicability. Therefore, this assay is expected to be useful for the control of forgery and alteration of raw materials in seafood.

목차 (Table of Contents)

• Ⅰ. 서 론 1
• Ⅱ. 재료 및 방법 5
• 1. 실험 재료 5
• 2. 실험 방법 8
• Ⅲ. 결과 및 고찰 25
• 1. 표준시료 유전학적 판별 25
• 2. 종 특이 primer 설계 26
• 3. Ultra-fast PCR 최적 반응조건 30
• 4. Ultra-fast PCR 정성 시험법 검증 33
• 5. Ultra-fast PCR 정량 시험법 검증 43
• 6. 모니터링 53
• Ⅳ. 요 약 58
• Ⅴ. 참고문헌 59