Although much smaller than base pairing, stacking interactions of the X(n+1)Y(n) type also make a substantial contribution to the duplex stability. In contrast, the effect of X(n)Y(n+1) type stacking is negligible as the corresponding bases are more distant due to the right-handed helical structure of DNA. We note that the magnitude of the base pairing compared with the stacking interactions aligns well with conventional textbook explanations, and with rupture force measurements obtained via atomic force microscopy (AFM). It is also worth mentioning here that interactions between distant pairs are negligible, thus lending fundamental theoretical support to popular nearest neighbor (NN) models, which assume no interaction between distant bases to predict thermodynamic data of any given DNA sequence. Overall, these results demonstrate that the fragment-pairwise LED scheme provides a theoretical framework that is uniquely suitable to disentangle the contribution of individual interactions to the stability of complex biomolecular assemblies, with applications across a myriad of fields of chemical research.