The production of fermentable sugars from renewable sources is a challenge. An attempt was made to exploit glycogen as a potential feedstock for the production of glucose. The microwave-assisted acidic hydrolysis was applied for glycogen decomposition for the first time. The optimal conditions for the hydrolysis reaction (yield of glucose – 62 wt.%) were identified: microwave irradiation time – 10 min and concentration of acid – 1 M HCl. Microwave irradiation has dramatically reduced the reaction time from more than 6 h (at 80 °C under an oil bath) to 10 min. ¹³C NMR spectroscopy was employed to monitor the progress of the hydrolysis reaction. HPLC analysis was employed to evaluate the yield of glucose. Thus, the viability of the use of glycogen as an economically and environmentally benign precursor to the production of glucose has been demonstrated.