Orientation changes in surface-bound hybridized DNA undergoing preparation for ex situ spectroscopic measurements

► The effect of rinsing on DNA hybrids in preparation for ex situ studies was examined. ► Light rinsing has a minor affect on the orientation and order of the hybrids. ► Strong rinsing disrupts the intactness and upright order in both target and probe DNA strands. ► These results demonstrate the importance of rinsing procedures for the preparation of hybridized DNA films for ex situ analysis.