The abnormal levels of four DNA bases, namely guanine (G), adenine (A), thymine (T), and cytosine (C) are implicated in several cancers, metabolic diseases, and HIV/AIDS. Therefore, the accurate detection and concentration measurement of these four DNA bases is of significant interest. Furthermore, there has recently been a push towards developing chemical sensors which are more sustainable and cost-effective. Herein, we developed a graphite paste electrode which incorporated the biochar of sugarcane and methylene blue (GPE-SC-MB) in order to simultaneously detect these four DNA bases. The linear ranges obtained for the four DNA bases are 0.67-38.67 mu M for G, 0.67-126.67 mu M for A, and 6.67-1600 mu M for T and C. The limit-of-detection (LOD) values obtained were 0.037 mu M for G, 0.042 mu M for A, 4.25 mu M for T, and 5.33 mu M for C. The electroactive surface area of the electrode as well as the diffusion coefficients for each analyte were determined. Lastly, the GPE-SC-MB was tested in real samples using human saliva with recovery values between 99.0 and 103.0%. Thus, biochar from sugarcane proved to be an effective electrode modifier material for the development of sensitive electrochemical sensors.