The invention provides a high-integration equidistance equipartition nucleic acid amplification micro-fluidic chip. The high-integration equidistance equipartition nucleic acid amplification micro-fluidic chip sequentially consists of a cover glass layer, a reaction layer, a decorative layer and a sealing layer from bottom to top in a thermal bonding manner, wherein the reaction layer is provided with n reaction modules, and each reaction module is provided with a circulation passage, a reaction micro-cavity and a sample feeding opening by utilizing a multilayer soft photoetching technology. By adopting the chip, the circulation distance of the sample before being introduced into a small chamber is identical, the sample feeding quantity is identical, and an error caused by the non-uniformity of the reaction volume can be avoided. The experiment cost is reduced. The sample feeding speed is increased. Quantitative detection in real time can be realized. The chip is reasonable in design, simple in structure, simple to operate and low in production cost and can be applied in the real-time fluorescent quantitative nucleic acid amplification, digital nucleic acid amplification, single-cell separation of nucleic acid amplification, SNP detection, detection of single-base mutation, detection of copy number imbalance, research of single-cell gene expression profile, early detection of cancer marker, stem cell differentiation identification, cell separation identification and the like.