The invention provides a manufacturing method of a semiconductor structure. The method includes: a) forming gate lines, which extend in a direction, on a substrate; b) forming a photoresist layer which covers the semiconductor structure and patterning the photoresist layer so as to form openings which cross the gate lines; c) forming self-assembled copolymers in the openings and reducing the openings; d) cutting off the gate lines through the openings so that the gate lines are insulated at the openings. Through forming add layers on the inner walls of the openings of the photoresist layer, a distance between two opposite walls of each opening in a gate width direction, that is, the distance between end parts of adjacent electrically isolated gates on the same line is reduced so that the area is saved and the integration degree of a semiconductor device is improved. Moreover, the invention also provides the semiconductor structure which is formed according to the method provided by the invention.