A method for patterning biomolecules using electron beam (e-beam) lithography has been developed. First, a non-biofouling poly (ethylene glycol) terminated self-assembled monolayer (SAM) was deposited on a silicon oxide substrate. This SAM was then ablated by e-beam to create patterns aligned with the pre-existing features on the wafer. Aldehyde-terminated polyamidoamine (PAMAM) dendrimers were assembled in a layer-by-layer fashion in the ablated patterns to allow the covalent immobilization of oligonucleotide probes. The aminated oligonucleotides were attached using Schiff base chemistry followed by reductive amination. The functionality of the attached oligonucleotides was demonstrated by the hybridization of fluorescently labeled complementary target oligonucleotides. The hybridized target oligonucleotides could be stripped and the regenerated surface bound probe oligonucleotides could be rehybridized with complementary target oligonucleotide.