<p>The development of a highly selective and sensitive Ba²⁺ sensor is crucial because of its industrial, environmental, and biological relevance. This study introduces a novel coated wire barium-selective electrode incorporating DNA as an ecofriendly natural ionophore in a plastic membrane, utilizing dioctyl phthalate as a plasticizer. The use of DNA as an ionophore provides enhanced selectivity and sensitivity, showcasing a slope of 33.15&#xa0;mV/decade across a broad concentration range (1 × 10⁻⁵ to 1 × 10⁻²&#xa0;M). The sensor exhibited a rapid response time of 9&#xa0;s, a wide pH tolerance (2.6–6.9), and good selectivity for Ba²⁺ over other cations. Characterization of the membrane using FT-IR, SEM, and EDX confirmed its structural and morphological features. Practical applicability was demonstrated by detecting Ba²⁺ in spiked samples (milk, juice, tap water and urine) with recovery rates of 96.07–98.9%. This DNA-based approach offers a promising advancement in ion-selective electrode technology, with significant implications for real-world Ba²⁺ detection.</p> Graphical Abstract <p></p>