Covalent attachment of single-stranded DNA to carbon paste electrode modified by activated carboxyl groups

A new method of the electrode modification and DNA immobilization for a biosensor is reported. Outer layer of a conventional carbon paste electrode (CPE) was modified with carboxyl groups by mixing stearic acid with the paste. Single-stranded deoxyribonucleic acid was attached to the modified electrode through a linker – ethylenediamine. Immobilization process was performed in the presence of activators – water soluble 1-ethyl-3(3′-dimethylaminopropyl)-carbodiimide (EDC) and N-hydroxysulfosuccinimide (NHS). Stearic acid concentration and other experimental parameters of the procedure were optimized. Covalent immobilization of DNA on the electrode surface exhibits some advantages as compared to simple adsorption mainly due to the fact that nucleic acid chains are bound to an electrode surface by one end only and it ensures structural flexibility and increases hybridization without DNA leakage. Modified electrode with immobilized (21-mer) oligonucleotide as a specific probe was successfully applied in preliminary investigations for the detection of bar gene commonly used in genetically modified food.