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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.

참고문헌 (22)

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이 논문을 인용한 문헌 (2)

  1. 김유영, 송인근, 민병례, 조홍범, 최영길 1999. "DNA 교잡에 의한 토양 미생물 군집의 다양성과 유사성" 환경생물 = Korean journal of environmental biology, 17(3): 279~284 
  2. 조재창 2001. "Environmental Genomics : Application of DNA Microarray Technology to Environmental Microbiology" 생물산업 : 한국미생물·생명공학회 소식지 = Bioindustry news, 14(2): 24~30 


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