Isolation of high molecular weight DNA from soil for cloning into BAC vectors

FEMS Microbiol Lett. 2003 Jun 6;223(1):15-20. doi: 10.1016/S0378-1097(03)00248-9.


Isolation of high molecular weight DNA fragments from soil, in excess of 1 Mb, and of sufficient quality for cloning into an Escherichia coli-streptomycete artificial chromosome vector is described. The combination of indirect extraction of cells, using a nycodenz extraction technique, followed by lysis of biomass immobilised in agarose plugs, allowed fragments in excess of 1 Mb to be purified.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacteria / genetics
  • Chromosomes, Artificial, Bacterial / genetics*
  • Cloning, Molecular
  • DNA, Bacterial / chemistry
  • DNA, Bacterial / genetics
  • DNA, Bacterial / isolation & purification*
  • Genetic Vectors
  • Italy
  • Molecular Weight
  • Soil Microbiology*
  • Trees


  • DNA, Bacterial