In this study, a new probe based on immobilization of amino linked oligonucleotide (NH2-linked DNA) on poly(glycidyl methacrylate-co-vinylferrocene)-coated Pt electrode was fabricated for the electrochemical detection of PTEN gene from human prostate tissues. The experimental parameters such as DNA immobilization time, DNA concentration, and target concentration were optimized. The selectivity of the NH 2-linked DNA probe was assessed with mismatch (MM) and noncomplementary (NC) sequences. The applicability of the NH2-linked DNA probe to the PCR amplified samples correspond to PTEN gene from prostate tissues was evaluated. The immobilization of DNA on the copolymer was confirmed by FTIR, AFM, CV and DPV analysis. The PCR products were also identified by using agarose gel electrophoresis. The prepared probe indicated a linear range (10-100 μg mL-1) with a detection limit (4.7 μg mL -1) and a good selectivity of the NH2-linked DNA probe toward target DNA sequence. © 2014 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2014, 131, 40638. Copyright © 2014 Wiley Periodicals, Inc.