Slalom chromatography (SC) is used for the separation of large double-stranded DNA molecules. In this technique, the progression of the DNA fragments through the closed column packing follows the flow direction and is like a snake edging is way into long grass. A novel mathematical model is developed in this paper to describe this hydrodynamic phenomenon. The results obtained provided a model for the resolution between two adjacent peaks on a chromatogram. As well, a chromatographic response function was used to obtain the most efficient separation conditions for a mixture of DNA fragments with sizes higher than 15 kbp in a minimum analysis time.