Figure 4

Amplified products after the tertiary round of PCR in genomic walking and T-DNA flanking sequence cloning using FPNI-PCR. (a) Cloning of the FT-Like gene of Rosa rugosa using 3 gene-specific and 1-8 FP primers. (b) T-DNA flanking sequence cloning in three transgenic tobacco individual plants using forward primers corresponding to the known T-DNA border; various fragment sizes were obtained from the use of different FP primers (1-9) in the three tested tobacco plants. (c) T-DNA flanking sequence cloning using backward primers. Only target specific fragments were amplified by using an appropriate annealing temperature during the low stringency cycles of the first PCR step (similar T-DNA sequences were present in each of the tested transgenic tobacco plants). Note: M: molecular marker; number: 1-9 FP primers; -: control.