Abstract. We report a simple method to isolate plasmids from single colonies of Agro bacterium tumefaciens by means of rolling circle amplification. The amplified DNA can be digested by restriction enzymes for plasmid verification and transformed into Escherichia coli for plasmid rescue. Compared with conventional procedures, this method eliminates liquid culturing of Agrobacterium cells and subsequent DNA isolation and enables large-scale plasmid analyses.
Key words: Agrobacterium tumefaciens, Escherichia coli, plasmid DNA, rolling circle amplification
Abbreviations: GUS, beta-glucuronidase; RCA, rolling circle amplification; TAP, tandem affinity purification.