Methods for conducting an assay to detect nucleic acid hybridization are
disclosed. In particular, the fabrication of a carbon electrode suited to
nucleic acid hybridization detection is described. In some preferred
embodiments, a micro array of carbon electrodes is constructed using
photolithography. The final step in the photolithography process involves
developing a solder mask with an alkaline solution. The alkaline solution
oxidizes the carbon surface producing surface carboxylic acid groups. The
surface carboxylic acid groups are reacted with EDC or DCC and NHS to
produce NHS esters. Immobilization of NEUTRAVIDIN.RTM. onto the electrode
surface is effected by reaction with the surface NHS ester groups. A
biotinylated probe DNA molecule is then attached to the electrode via
binding between the biotin group and the immobilized NEUTRAVIDIN.RTM..
The resulting electrode can be used to detect hybridization between the
probe DNA and complementary target DNA.