Nucleic acid extraction from Agrobacterium strains.
Agrobacterium is routinely used as a tool for moving genetic constructs into plant cells. The successful use of Agrobacterium as a tool for the genetic engineering of plant cells often requires the manipulation and analysis of nucleic acids present in recombinant Agrobacterium strains. Here we present dependable methods for the isolation of genomic (total) DNA, mega-plasmid DNA, shuttle or binary plasmid DNA, and RNA. In addition, we provide a simple method for the electronic transfer of shuttle plasmids from Agrobacterium to E. coli for use when their low copy number in Agrobacterium impedes plasmid isolation from that strain.
Wise
Arlene A
AA
Plant Science Institute, Department of Biology, University of Pennsylvania, PA, USA.
Liu
Zhenying
Z
Binns
Andrew N
AN
D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification*; D018274:Electroporation / Q000379:methods; D004926:Escherichia coli / Q000737:chemistry* / Q000235:genetics / Q000254:growth & development; D059828:Plant Cells; D036741:Plant Tumor-Inducing Plasmids / Q000235:genetics / Q000302:isolation & purification*; D010944:Plants / Q000235:genetics / Q000382:microbiology; D012329:RNA, Bacterial / Q000235:genetics / Q000302:isolation & purification*; D012231:Rhizobium / Q000737:chemistry* / Q000235:genetics / Q000254:growth & development; D013045:Species Specificity
D004269:DNA, Bacterial; D012329:RNA, Bacterial