The sequence method was first described by Di Rienzo in cats and applied in different species including humans. Until now, no systematic study of spontaneous baroreflex sensitivity (BRS) has been performed by the sequence method in mice. This study aimed to characterize the best estimates of BRS using the sequence method by tuning all the possible parameters, specifically, the number of beats involved in a sequence, the minimal changes in blood-pressure (BP) ramps, and the minimal changes in pulse-interval (PI) ramps. Also, the relevance to set a minimal correlation coefficient in the regression line between BP and PI was tested. An important point was the delay to be applied between BP and PI. This delay represents the physiological time for the baroreflex loop to efficiently correct the BP variations.