1.5

1. Kb 10 5 3 1.5 1 Figure 1 - Supplementary Hind III Eco RV Figure 1 –PdKn1 gene copy number. Southern blot analysis for PdKn1 using as probe the internal sequence of the gene (amino acid region b-d in Figure 2A) that contains two very conserved motives in Knox genes. Eco RV cuts once inside the probe template. For PdKn1 Southern blot analysis Eco RV and Hind III restriction enzymes were used separately to digest 10mg aliquots of genomic DNA. Digested DNA was electrophoresed in a 0.8% 1xTAE agarose/EtBr gel, run overnight at 2V/cm and blotted overnight onto a Hybond+ nylon membrane. The hybridisation was performed overnight at 61C, and the stringency washes, the first (1xSSC and 0.1% SDS) and second (0.5xSSC and 0.1% SDS), were run at 61C for 45 and 20 min, respectively. The membrane was exposed to a Kodak film for 1h at room temperature, in the dark. Digestion with Hind III reveals at least 5 bands