Recovery, purification, and cloning of high-molecular-weight DNA from soil microorganisms

Appl Environ Microbiol. 2008 May;74(10):3302-5. doi: 10.1128/AEM.02630-07. Epub 2008 Mar 21.

Abstract

We describe here an improved method for isolating, purifying, and cloning DNA from diverse soil microbiota. Soil microorganisms were extracted from soils and embedded and lysed within an agarose plug. Nucleases that copurified with the metagenomic DNA were removed by incubating plugs with a high-salt and -formamide solution. This method was used to construct large-insert soil metagenomic libraries.

MeSH terms

  • Cloning, Molecular*
  • DNA / genetics*
  • DNA / isolation & purification*
  • Deoxyribonucleases / antagonists & inhibitors
  • Enzyme Inhibitors / pharmacology
  • Formamides / pharmacology
  • Molecular Biology / methods*
  • Molecular Weight
  • Soil Microbiology*

Substances

  • Enzyme Inhibitors
  • Formamides
  • formamide
  • DNA
  • Deoxyribonucleases