Allowance for heterogeneous stacking in the DNA helix-coil transition theory.

Melting profiles of eight DNA molecules with lengths ranging from 849 to 4362 bp have been measured in an SSC buffer where the melting is an equilibrium process up to complete strand separation. A theoretical analysis shows that the melting profiles depend on only eight invariants that are linear combinations of 10 original stacking parameters. As a result it is impossible to determine the 10 parameters from the melting profiles. The 8 variants have been determined by fitting theoretical profiles to experimental ones for two fragments. Then theoretical and experimental profiles are compared for those 6 fragments that were not used in the fitting procedure. This comparison demonstrates that allowance for heterogeneous stacking considerably improves the agreement between theory and experiment. The values of invariants have proved to be small. This confirms the previous conclusion that heterogeneous stacking interactions produce only small corrections to the major effect of the difference in the mean stabilities of AT and GC pairs. Some discrepancy between theory and experiment that remains after the allowance for heterogeneous stacking is probably due to even finer effects of long-range interactions.