A detection chip, a method of using a detection chip and a reaction system are provided. The detection chip includes a first substrate, a micro-chamber definition layer and a heating electrode. The micro-chamber definition layer is located on the first substrate and defines a plurality of micro-reaction chambers. The heating electrode is located on the first substrate and closer to the first substrate than the micro-chamber definition layer, and configured to release heat after being energized. The heating electrode includes a plurality of sub-electrodes, orthographic projections of the plurality of micro-reaction chambers on the first substrate overlap with orthographic projections of at least two of the plurality of sub-electrodes on the first substrate, and the at least two of the plurality of sub-electrodes have different heating values per unit time after being energized.