An ambiguity fixing method based on sequential least squares of the present invention can improve the success rate of measuring the ambiguity between two antennas in real time by using the single-frequency or double-frequency navigation satellite signals in a dynamic environment, provides the data support for the subsequent high-precision baseline measurement, attitude determination and the like,and is characterized by introducing the priori information in the ambiguity real number solution estimation process to serve as the state information to be transmitted to a filter, and accelerating convergence while reducing the morbidity of a design matrix. According to the method, the thought based on a measurement error modeling theory and a matrix triangulation decomposition technology is applied under a sequential condition, the additional calculation burden is not increased while the measurement information is accumulated, an ambiguity real number solution and the cofactor matrix information are calculated by constructing an ambiguity filter in a least square form, and finally, the ambiguity real number solution information is processed by using an LAMBDA method to search an ambiguity integer solution, thereby solving the problem of low ambiguity solving success rate of navigation satellite signals in a dynamic environment in a traditional method.