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A formula for thermal stability (Tm) prediction of PNA/DNA duplexes
U Giesen1, W Kleider, C Berding
1Boehringer Mannheim GmbH, Roche Diagnostics, Business Unit Laboratory Systems, Bahnhofstr. 9-15,82327 Tutzing, Germany.
Abstract:
An empirical formula for thermal stability (T m) prediction of PNA/DNA duplexes has been derived. The model is based on the T m as calculated for the corresponding DNA/DNA duplex employing a nearest neighbour approach, by including terms for the pyrimidine content and length of the PNA to take into account the increased thermostability of PNA/DNA hybrids and the asymmetry of the PNA-DNA heteroduplex. The predictive power of the T m prediction formula was challenged with an independent data set not used for model building. The T m of >90% of the sequences was predicted within 5 K; 98% of the predicted T ms differ by not more than 10 K from the experimentally determined T m.