Fig. 2

Abl kinase domain T315I point mutation detection analysis by PNA directed PCR clamping. A representative result of the analysis carried out on cDNAs isolated from patients affected by imatinib resistance CML is represented in panel (a). PCR amplification was carried-out in absence (−) or in presence (+) of competitor PNA, at a concentration 3× greater than primer FWD. The amplification performed without (−) PNA represents an internal positive control displaying the efficiency of template amplification. As result only when PNA-template duplex stability is weakened because of the mutation an efficient template amplification occurred. L: DNA ladder. Sensitivity was assessed mixing, at different ratio, mutated (T315I) and w.t. template panel (b). Dilutions were as follow: 100, 20, 10, 5, 1, 0.5 and 0 % mutated (T315I) versus w.t. template