The invention relates to a filling method of a large-area zinc electrolytic negative plate pre-stripping port, which can realize the batch and scale production of the filling of the negative plate pre-stripping port. The method has the characteristics of low cost, simplicity, convenience, convenience in operation and easiness for controlling the product quality. In the method, a pre-stripping port filling mold, a clamp, an insertion plate groove, a pneumatic glue gun (containing a mixing nozzle) and an air compressor are mainly adopted. The filling method has the following remarkable characteristics that (1) the mold is low in cost and convenient to maintain; (2) the production labor intensity is low; (3) the land can be saved; (4) the batch and scale production can be realized; (5) the rate of pre-stripping port finished products is high.