Abstract
We report a simple method to isolate plasmids from single colonies ofAgrobacterium tumefaciens by means of rolling circle amplification. The amplified DNA can be digested by restriction enzymes for plasmid verification and transformed intoEscherichia coli for plasmid rescue. Compared with conventional procedures, this method eliminates liquid culturing ofAgrobacterium cells and subsequent DNA isolation and enables large-scale plasmid analyses.
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Abbreviations
- GUS:
-
beta-glucuronidase
- RCA:
-
rolling circle amplification
- TAP:
-
tandem affinity purification
References
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Chen, X., Ding, X. & Song, WY. Isolation of Plasmid DNA rescued from single colonies ofAgrobacterium tumefaciens by means of rolling circle amplification. Plant Mol Biol Rep 21, 411–415 (2003). https://doi.org/10.1007/BF02772590
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DOI: https://doi.org/10.1007/BF02772590