• Title/Summary/Keyword: ERIC-PCR components

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Optimization of the Concentrations of ERIC-PCR Components to Simultaneously Differentiate Five Foodborne Pathogenic Bacterial Genera (식중독세균 5속의 동시 동정을 위한 ERIC-PCR 반응성분 농도의 최적화)

  • Seo, Hyun-Ah;Park, Sung-Hee;Kim, Keun-Sung
    • Journal of Food Hygiene and Safety
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    • v.18 no.4
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    • pp.229-236
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    • 2003
  • The five different foodborne pathogenic bacterial genera of Escherichi, Salmonella, Shigella, Vibrio and Listeria are important sources of foodpoison. However, the method was not developed to simultaneously differentiate these five bacteria at molecular level. The optimized concentrations of the four major PCR cocktail components of $MgCl_2$, dNTPs, primers and template DNA were determined when ERIC (enterobacterial repetitive intergenic consensus)-PCR reactions were carried out to differentiate the five differnet foodborne pathogenic bacteria. The optimized concentration of $MgCl_2$ was determined to be 2 mM in order to obtain a consistent fingerprinitng pattern. The similar fingerprinting pattern was obtained when ERIC primers and dNTPs were added up to the concentrations of 2 ${\mu}M$ and 200 ${\mu}M$, respectively. As for template DNA, the numbers of PCR fragments were not affected, but their intensities were increased as the concentrations of the DNA were increased.