본문 바로가기
HOME> 논문 > 논문 검색상세

논문 상세정보

The journal of microbiology v.34 no.3, 1996년, pp.229 - 235   피인용횟수: 2

Direct Extraction of DNA from Soil for Amplification of 16S rRNA Gene Sequences by Polymerase Chain Reaction

Cho, Jae-Chang    (Research Center for Molecular Microbiology, Seoul National University   ); Lee, Dong-Hun    (Research Center for Molecular Microbiology, Seoul National University   ); Cheol, Cho-Young    (Research Center for Molecular Microbiology, Seoul National University   ); Cho, Jang-Cheon    (Research Center for Molecular Microbiology, Seoul National University ;  ); Kim, Sang-Jong  
  • 초록

    Microgram quantities of DNA per gram soil were recovered with SDS- based and freeze-and thaw procedures. The average DNA fragment size was > 23 Kb. This method generated minimal shearing of extracted DNA. However, the DNA extracts still contained considerable amounts of humic impurities sufficient to inhibit PCR. Several approaches were used to reduce the interferences with the PCR (use of CTAF in extraction step, Elutip-d column purification, addition of BSA to PCR buffer) to accomplish PCR with DNA extract as a template. Most of the DNA extracts were not digested completely by restriction endonuclease, and CTAB-TREATED ane Elutip-d column purified DNA extracts were partially digested. Regarding as restriction enzyme digestion, all PCRs failed to amplify 16S rRNA gene fragments in the DNA extracts. In the case of DNA extracts only where BSA was added to PCR buffer, PCR was successfully conducted whether the DNA extracts were treated with CTAB or purified with columns. However, these two treatments were indispensable for humic impurity-rich DNA extracts to generate the PCR-compatible DNA samples. Direct extraction of DNA, coupled with these procedures to remove and relieve interferences by humic impurities and followed by the PCR, can be rapid and simple method for molecular microbiological study on soil microorganisms.


  • 주제어

    Soil .   DNA extraction .   PCR.  

  • 참고문헌 (22)

    1. Bacterial and fungal activities in soil: separation of bacteria and fungi by a rapid fractionated centrifugation technique , Fagri, A.;V.L. Torsviik;J. Goks yr , Soil Biol. Biochem. / v.9,pp.105-112,
    2. Extraction from natural planktonic microorganisms of DNA suitable fro molecular biological studies , Fuhrman, J.A.;D.E. Comeau;A. Hagstrom;A.M. Chan , Appl. Environ. Microbiol. / v.54,pp.1426-1429,
    3. DNA probe method for the detection of specific microorganisms in the soil bacterial community , Holben, W.E,;J.K. Jansson;B.K. Chelm;J.M. Tiedje , Appl. Environ. Microbiol. / v.54,pp.703-711,
    4. Ingraham, J.L.;O. Maaloe;F.C. Neidhardt , Growth of the bacterial cell / v.,pp.1-48,
    5. Survival strategies of bacteria in the natural environment , Roszak, D.B.;R.R. Colwell , Microbiol. Rev. / v.51,pp.365-379,
    6. Simple, rapid method for direct isolation of nucleic acids from aquatic environments , Somerville, C.C.;I.T. Knight;W.L. Straube;R.R. Colwell , Appl. Environ. Microbiol. / v.55,pp.548-554,
    7. Recovery of DnA from soils and sediments , Steffan, R.J.;J. GoksoyrlA.K. Bej;R.M. Atlas , Appl. Environ. Microbiol. / v.54,pp.2908-2915,
    8. DNA recovery from soils of diverse composition , Zhou, J.;M.A. Bruns;J.M. Tiedje , Appl. Environ. Microbiol. / v.62,pp.316-322,
    9. Molecular ecological study on the structure of bacterial communities in lake soyang , Lee, D.-H. , Ph. D. thesis, Seoul National University / v.,pp.,
    10. Isolation of bacterial DNA from soil , Torsvik, V.L. , Soil Biol. Biochem. / v.12,pp.15-21,
    11. Sambroock, J.;E.F. Fritsch;T. Maniatis , Molecular cloning : a laboratory manual, (2nd ed) / v.,pp.,
    12. ASA-SSSA , Methods of Soil Analysis part2. / v.,pp.,
    13. Simplified method for dissolved DNA determination in aquatic environments , DeFlaun, M.F.;J.H. Paul;D. Davis , Appl. Environ. Microbiol. / v.52,pp.654-659,
    14. Overcoming problems of phenolics and quinones in the isolation of plant enzymes and organelles , Loomis, W.D. , Methods Enzymol / v.31,pp.528-545,
    15. Fluorometric determination of DNA in aquatic microorganisms by use of Hoechst 33258 , Paul, J.H.;B. Myers , Appl. Environ. Microbiol. / v.43,pp.1393-1399,
    16. Detection of polychlorinated biphenyl degradation genes in polluted sediment by direct DNA extraction and polymerase chain reaction , Erb, R.W.;I. Wagner-Dovler , Appl. Environ. Microbiol. / v.59,pp.4065-4073,
    17. The extraction and purification of microbial DNA from sediments , Ogram, A.;G.S. Sayler;T. Barkay , J. Microbiol. Methods / v.7,pp.57-66,
    18. Proteinase inhibition of the detection of Listeria monocytogenes in milk using the PCR , Powell, H.A.;C.M. Gooding;S.D. Garreff;B.M. Lund;R.A. McKee , Lett. Appl. Microbiol. / v.18,pp.59-61,
    19. Relief of amplication inhibition in PCR with bovine serum albumin or T4 gene 32 protein , Kreader, C.A. , Appl. Environ. Microbiol. / v.62,pp.1102-1106,
    20. DNA extraction and purification from sediments , Ogram, A.;G.S. Sayler;T. Barkay , J. Microbiol. Methods / v.7,pp.57-66,
    21. Rapid method for direct extraction of DNA from soil and sediments , Tsai, Y.L.;B.H. Olson , Appl. Environ. Microbiol. / v.57,pp.1070-1074,
    22. Rapid method for separation of bacterial DNA from humic substances in sediments for polymerase chain reaction , Tsai, Y.L.;B.H. Olson , Appl. Environ. Microbiol. / v.58,pp.2292-2295,
  • 이 논문을 인용한 문헌 (2)

    1. 1999. "The Diversity and Similarity of Soil Microbial Communities by DNA Cross Hybrization" 환경생물 = Korean journal of environmental biology, 17(3): 279~284     
    2. 2001. "" 생물산업 : 한국미생물·생명공학회 소식지 = Bioindustry news, 14(2): 24~30     

 활용도 분석

  • 상세보기

    amChart 영역
  • 원문보기

    amChart 영역

원문보기

무료다운로드
  • NDSL :
유료다운로드

유료 다운로드의 경우 해당 사이트의 정책에 따라 신규 회원가입, 로그인, 유료 구매 등이 필요할 수 있습니다. 해당 사이트에서 발생하는 귀하의 모든 정보활동은 NDSL의 서비스 정책과 무관합니다.

원문복사신청을 하시면, 일부 해외 인쇄학술지의 경우 외국학술지지원센터(FRIC)에서
무료 원문복사 서비스를 제공합니다.

NDSL에서는 해당 원문을 복사서비스하고 있습니다. 위의 원문복사신청 또는 장바구니 담기를 통하여 원문복사서비스 이용이 가능합니다.

이 논문과 함께 출판된 논문 + 더보기