A delay line circuit with a calibration function, includes N delay modules and a calibration module. The N delay modules are serially coupled to each other. The calibration module generates a calibration start signal and a calibration stop signal according to a calibration signal and a clock signal, and the calibration start signal is outputted to the N delay modules, so that the N delay modules output N delay signals according to N control signals and the calibration start signal. The calibration module calibrates the N control signals according to the N delay signals and the calibration stop signal, so that the N delay modules generate N calibrated delay signals according to the N calibrated control signals and the clock signal. A generation time instant of the calibration stop signal is later than a generation time instant of the calibration start signal.