title: Recovery, purification, and cloning of high-molecular-weight DNA from soil microorganisms.
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.
authors: Department of Biological Sciences, Auburn University, Auburn, Alabama, USA.
authors: Mark R
authors: MR
authors: Liles
authors: Lynn L
authors: LL
authors: Williamson
authors: Jitsupang
authors: J
authors: Rodbumrer
authors: Vigdis
authors: V
authors: Torsvik
authors: Robert M
authors: RM
authors: Goodman
authors: Jo
authors: J
authors: Handelsman
mesh_terms: D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics* / Q000302:isolation & purification*; D003851:Deoxyribonucleases / Q000037:antagonists & inhibitors; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005559:Formamides / Q000494:pharmacology; D008967:Molecular Biology / Q000379:methods*; D008970:Molecular Weight; D012988:Soil Microbiology
chemical_list: D004791:Enzyme Inhibitors; D005559:Formamides; C031066:formamide; D004247:DNA; D003851:Deoxyribonucleases