Isolation of polymerase chain reaction-ready bacterial DNA from Lake Baikal sediments by carboxyl-functionalised magnetic polymer microspheres

J Chromatogr A. 2006 Oct 13;1130(1):115-21. doi: 10.1016/j.chroma.2006.07.034. Epub 2006 Aug 9.

Abstract

Carboxyl group-containing magnetic nonporous poly(2-hydroxyethyl methacrylate-co-ethylene dimethacrylate) (P(HEMA-co-EDMA)) microspheres were used for the isolation of polymerase chain reaction (PCR)-ready DNA from samples of Baikal sediments. DNA was isolated using the phenol extraction method or the Soil DNA Isolation Kit. The occurrence of false-negative results in PCR caused by the presence of extracellular inhibitors in DNA samples was solved using solid phase reversible DNA immobilisation. PCR-ready DNA was reversibly adsorbed to the microspheres in the presence of 8.0% (w/v) poly(ethylene glycol) (PEG 6000) and 2.0M sodium chloride concentrations. The adsorbed DNA was released from the microspheres in a low ionic strength TE buffer. The quality of isolated DNA was checked by PCR amplification.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacillus / chemistry
  • DNA, Bacterial / chemistry
  • DNA, Bacterial / isolation & purification*
  • Electrophoresis, Agar Gel
  • Fresh Water / analysis
  • Geologic Sediments / analysis*
  • Magnetics*
  • Microspheres*
  • Polymerase Chain Reaction
  • Polymethacrylic Acids / chemistry*

Substances

  • DNA, Bacterial
  • Polymethacrylic Acids
  • poly(2-hydroxyethyl methacrylate-co-ethylene dimethacrylate)