Isolation of high molecular weight DNA from soil for cloning into BAC vectors.
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.
Berry
Andrew E
AE
Department of Biological Sciences, University of Warwick, Gibbet Hill Rd, Coventry CV4 7AL, UK. aberry@bio.warwick.ac.uk
Chiocchini
Claudia
C
Selby
Tina
T
Sosio
Margherita
M
Wellington
Elizabeth M H
EM
D001419:Bacteria / Q000235:genetics; D022202:Chromosomes, Artificial, Bacterial / Q000235:genetics*; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D005822:Genetic Vectors; D007558:Italy; D008970:Molecular Weight; D012988:Soil Microbiology; D014197:Trees
D004269:DNA, Bacterial