Plant Molecular Biology

, Volume 17, Issue 1, pp 151–153

A quick method to estimate the T-DNA copy number in transgenic plants at an early stage after transformation, using inverse PCR

  • Mirjam P. Does
  • Ben M. M. Dekker
  • Marcel J. A. de Groot
  • Remko Offringa
Update Section Novel Method

Abstract

Agrobacterium-mediated transformation of plants is known to result in transgenic plants with a variable number of integrated T-DNA copies [1, 2, 3, 7]. Our aim was to obtain transgenic tobacco plants containing one integrated T-DNA copy per genome. Therefore, a quick method was developed to estimate the T-DNA copy number of young transgenic plantlets within 10 weeks after transformation. Inverse polymerase chain reaction (IPCR) was used to amplify junction fragments, i.e. plant genomic DNA sequences flanking the known T-DNA sequences [5].

Key words

Agrobacterium-mediated transformation inverse PCR Nicotiana tabacum SRI T-DNA copy number 

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Copyright information

© Kluwer Academic Publishers 1991

Authors and Affiliations

  • Mirjam P. Does
    • 1
  • Ben M. M. Dekker
    • 1
  • Marcel J. A. de Groot
    • 1
  • Remko Offringa
    • 1
  1. 1.MOGEN International nvLeidenNetherlands

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