Nucleic Acids Research, Vol 27, Issue 24 4801-4806, Copyright © 1999 by Oxford University Press
MC Chakrabarti and FP Schwarz
Thermodynamics of the thermal dissociation transitions of 10 bp PNA/DNA
duplexes and their corresponding DNA/DNA duplexes in 10 mM sodium phosphate
buffer (pH 7.0) were determined from differential scanning calorimetry
(DSC) measurements. The PNA/DNA transition temperatures ranged from 329 to
343 K and the calorimetric transition enthalpies ranged from 209 +/- 6 to
283 +/- 37 kJ mol(-1). The corresponding DNA/DNA transition temperatures
were 7-20 K lower and the transition enthalpies ranged from 72 +/- 29 to
236 +/- 24 kJ mol(-1). Agreement between the DSC and UV monitored melting
(UVM) determined transition enthalpies validated analyzing the UVM
transitions in terms of a two- state transition model. The transitions
exhibited reversibility and were analyzed in terms of an AB = A + B
two-state transition model which yielded van't Hoff enthalpies in agreement
with the transition enthalpies. Extrapolation of the transition enthalpies
and free energy changes to ambient temperatures yielded more negative
values than those determined directly from isothermal titration calorimetry
measurements on formation of the duplexes. This discrepancy was attributed
to thermodynamic differences in the single-strand structures at ambient and
at the transition temperatures, as indicated by UVM measurements on single
DNA and PNA strands.
ARTICLES
Thermal stability of PNA/DNA and DNA/DNA duplexes by differential scanning calorimetry
Center for Advanced Research in Biotechnology/National Institute of Standards and Technology, 9600 Gudelsky Drive, Rockville, MD 20850, USA.
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