Efficient Removal of Humic Substances in Preparing DNA Extract from Marine Sediments

해양퇴적물의 핵산추출물에서 humic substances의 효율적인 제거방법

  • Lee, Jung-Hyun (Marine Microbiology Lab., Korea Ocean Research & Development Institute) ;
  • Shin, Hyun-Hee (Marine Microbiology Lab., Korea Ocean Research & Development Institute) ;
  • Lee, HongKun (Marine Microbiology Lab., Korea Ocean Research & Development Institute) ;
  • Kwon, Kae Kyoung (Marine Microbiology Lab., Korea Ocean Research & Development Institute) ;
  • Ki, Sang-Jin (Marine Microbiology Lab., Korea Ocean Research & Development Institute)
  • 이정현 (한국해양연구소 생물부 미생물연구실) ;
  • 신현희 (한국해양연구소 생물부 미생물연구실) ;
  • 이홍금 (한국해양연구소 생물부 미생물연구실) ;
  • 권개경 (한국해양연구소 생물부 미생물연구실) ;
  • 김상진 (한국해양연구소 생물부 미생물연구실)
  • Received : 1998.04.30
  • Accepted : 1998.06.10
  • Published : 1998.09.01

Abstract

Molecular analyses of natural microbial communities are often dependent upon the obtainments of pure nucleic acids. The four methods (elution after agarose gel electrophoresis, G-75 microspin columns, hydroxyapatite mi-crospin columns, and polyvinylpolypyrrolidone (PVPP) microspin columns) were compared for the removal of PCR-inhibitory humic substances from the crude DNA extracts of marine sediment samples. The PVPP microspin columns have shown superior removal of humic substances from the crude DNA extract of marine sediment samples, with yield of $4.8{\mu}g/g$ (dry weight of sediment). The purified DNA by this rapid method was pure enough to amplify 1.5 kb fragment corresponding almost full length of 16S rRNA genes.

Keywords

DNA extraction;humic subsances;marine sediment;16S rRNA

Acknowledgement

Supported by : 한국해양연구소