The long-range film structure of 15 base pair, thiol-derivatized duplexes t
ethered through single 3 ' and 5 ' linkages to a Au(111) surface is measure
d via scanning probe microscopy. Film morphology of the 3 ' linked duplexes
differs from that of its 5 ' counterpart as observed by scanning probe mic
roscopy in contact mode under buffer solution. No structural features are d
istinguished in a film formed with the 5 ' linked duplexes as measured with
either a bare silicon nitride or chemically modified probe. However, a dis
tinct pattern in the film structure of the 3 ' linked duplexes is measured
with a chemically modified, hydrophobic tip. Small changes in linker orient
ation can greatly affect DNA film structure, and a hypothesis for the self-
assembly mechanism of the 3 ' linked duplexes is presented. Factors such as
linker placement and composition must be considered in comparing DNA micro
arrays formed with different tethering methods.