Ribociclib (RIB) is a cyclin-dependent kinase inhibitor used in the treatment of breast cancer. In this study, for the first time, the electrochemical behavior, quantification and interaction of RIB with DNA were carried out using voltammetric, spectrophotometric and molecular docking techniques. An environmentally-friendly disposable pencil graphite electrode (PGE) sensor was used as the working electrode. Utilizing the cyclic voltammetry technique, RIB produced an irreversible anodic wave around +0.837 V and reversible signals around +0.278 V/+0.209 V on the PGE surface. Using the PGE sensor, the RIB compound gave a very distinct anodic signal at a potential of +0.77 V in PBS (pH 3.0). For this analytical signal, the limit of detection and limit of quantitation values of RIB in the concentration range of 0.0139 3 M–0.0973 3 M in PBS (pH 3.0) were determined as 2.84 nM and 9.46 nM, respectively. The interaction of RIB with DNA was studied by voltammetric, spectrophotometric and molecular docking techniques. In the evaluation of the results obtained with all three techniques, the interaction of the RIB molecule with DNA was determined to occur through minor groove binding.