The ionic liquid (IL) modified chemically activated (CA) pencil graphite electrodes (PGEs) were developed for label-free voltammetric detection of miRNA-34a, and implemented to the real samples. Firstly, the electrochemical characterization of unmodified PGE, CA-PGE, IL-PGE and IL-CA-PGE was performed by cyclic voltammetry (CV) as well as their DNA binding capacity was studied by electrochemical impedance spectroscopy (EIS) technique. The microscopic characterization of the surface of each electrodes was investigated by scanning electron microscopy (SEM). Differential pulse voltammetry (DPV) technique was used for measuring the oxidation signal of guanine in order to perform a label-free voltammetric monitoring of a full-match hybridization specific to miRNA-34a. The selectivity of biosensor was tested against to miRNA-155, miRNA-660 as well as to the mismatch sequence of miRNA-34a. The further selectivity of this proposed biosensor was studied in the mixture of samples containing miRNA-34a with other miRNAs (1 : 1). The voltammetric detection of miRNA-34a was also explored in the artificial serum medium as fetal bovine serum (FBS) and also in total RNA samples isolated from HUH-7 human hepatocellular carcinoma cell line.