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A quick method to estimate the T-DNA copy number in transgenic plants at an early stage after transformation,using inverse PCR
Authors:Mirjam P Does  Ben M M Dekker  Marcel J A de Groot  Remko Offringa
Institution:(1) MOGEN International nv, Einsteinweg 97, 2333 CB Leiden, Netherlands
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].
Keywords:Agrobacterium-mediated transformation  inverse PCR  Nicotiana tabacum SRI  T-DNA copy number
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