• Title/Summary/Keyword: SYBR green

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Development and Verification of and Single Nucleotide Polymorphism Markers toDetermine Country of Origin of Korean and Chinese Scapharca subcrenata (한국산과 중국산 새꼬막(Scapharca subcrenata)의 원산지 판별을 위한 SNP 마커의 개발 및 검증)

  • Seong Seok Choi;Seung Hyun Yoo;Yong Bae Seo;Jong Oh Kim;Ik Jung Kwon;So Hee Bae;Gun Do Kim
    • Journal of Life Science
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    • v.33 no.12
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    • pp.1025-1035
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    • 2023
  • In this study, we analyzed SNPs that appear between Korean and Chinese Scapharca subcrenata using the nucleotide sequence data of S. subcrenata analyzed by genotyping by sequencing (GBS). To distinguish the country of origin for S. subcrenata in Korean and Chinese, we developed a primer set as single nucleotide polymorphism (SNP) markers for quantitative real-time PCR (qPCR) analysis and validated by sequencing SNPs. A total of 180 samples of S. subcrenata were analyzed by genotyping by sequencing, and 15 candidate SNPs were selected. SNP marker selection for country of origin were identified through real-time qPCR. Insertion 1 and SNP 21 markers showed the most distinct separation between the sequence types as well as the country of origin through qPCR, with the observed amplification patterns matching the expected outcomes.. Additionally, in a blind test conducted by mixing samples of S. subcrenata at random, Insertion 1 showed 74% accuracy, 52% sensitivity, and 96% specificity, and SNP 21 showed 86% accuracy, 79% sensitivity, and 93% specificity. Therefore, the two SNP markers developed are expected to be useful in verifying the authenticity of the country of origin of S. subcrenata when used independently or in combination.