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Isolation of Plasmid DNA rescued from single colonies ofAgrobacterium tumefaciens by means of rolling circle amplification

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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

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Authors and Affiliations

  1. Department of Plant Pathology, Plant Molecular and Cellular Biology Program, University of Florida, 32611, Gainesville, FL, USA

    Xiuhua Chen, Xiaodong Ding & Wen-Yuan Song

Authors
  1. Xiuhua Chen
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  2. Xiaodong Ding
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  3. Wen-Yuan Song
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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

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