Recovery, purification, and cloning of high-molecular-weight DNA from soil microorganisms.
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.
Liles
Mark R
MR
Department of Biological Sciences, Auburn University, Auburn, Alabama, USA.
Williamson
Lynn L
LL
Rodbumrer
Jitsupang
J
Torsvik
Vigdis
V
Goodman
Robert M
RM
Handelsman
Jo
J
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
D004791:Enzyme Inhibitors; D005559:Formamides; C031066:formamide; D004247:DNA; D003851:Deoxyribonucleases