The Korean Journal of Mycology (한국균학회지)

The Korean Society of Mycology (한국균학회)
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Parameters Affecting Polymerase Chain Reaction in RAPD Analysis of Pleurotus spp.

느타리버섯속(屬)의 DNA 다형성분석(多型性分析)에 영향(影響)을 미치는 PCR 조건(條件)

  • Kim, Beom-Gi (Division of Molecular Genetics, National Agricultural Science and Technology Institute, RDA) ;
  • Jeong, Mi-Jeong (Division of Molecular Genetics, National Agricultural Science and Technology Institute, RDA) ;
  • Lee, Chang-Soo (Division of Molecular Genetics, National Agricultural Science and Technology Institute, RDA) ;
  • Lee, Hee-Kyung (Division of Molecular Genetics, National Agricultural Science and Technology Institute, RDA) ;
  • Yoo, Young-Bok (Division of Molecular Genetics, National Agricultural Science and Technology Institute, RDA) ;
  • Ryu, Jin-Chang (Division of Molecular Genetics, National Agricultural Science and Technology Institute, RDA)
  • 김범기 (농업과학기술원 분자유전과) ;
  • 정미정 (농업과학기술원 분자유전과) ;
  • 이창수 (농업과학기술원 분자유전과) ;
  • 이희경 (농업과학기술원 분자유전과) ;
  • 유영복 (농업과학기술원 분자유전과) ;
  • 유진창 (농업과학기술원 분자유전과)
  • Published : 1995.09.30
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Abstract

This study describes the effects of several components on PCR amplification used for RAPD. We used different concentrations of reaction components to obtaine discrete and reproducible PCR products from Pleurotus cornucopiae. The optimum concentrations of reaction components were found to be 80 ng of template DNA, 30 pmole of 10-mer primer, $200\;{\mu}M$ dNTP, 2mM $MgCl_2$, 50 mM KCl, 10 mM Tris-HCl(pH 9.0), 0.1% Triton X-100, 1.5 unit of Taq DNA polymerase (promega) in $50\;{\mu}l$ reaction volume. The optimum annealing temperature was $35^{\circ}C$. These results proved to be valuable for characterization of Pleurotus spp.

본 실험결과 느타리버섯속에서의 재현성 있는 최적 RAPD 조건은 $50\;{\mu}l$ 반응액에서 80 ng template DNA, 30 pmole primer, $200\;{\mu}M$ dNTP, 2 mM $MgCl_2$, 50 mM KCl, 10 mM Tris-HCI(pH 9.0), 0.1% Triton X-100, 1.5 unit Taq polymerase(promega)였다. 여섯 개의 primer가 Pleurotus속 8종의 균주에서 RAPD polymorphism을 보임을 알 수 있었으며, 이들의 염기배열은 PR2(GGG GGG AAG C), PR3(GCG GTT GAG G), PR4(CGC ACC GCA C), PR10(CAA TCG CCG T), PR11(CAG CAC CCA C), PR17(TAG GCG TAT CAG GAG GCC CT)이었다.

Keywords

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