This manuscript aims to study the reliability of different variables related to performance and acceleration during the golf putt in players with medium-to-high handicaps and to determine the number of attempts necessary to find reliable values for these variables. Eight males and two females [55.67 (13.64) years, 78.4 (11.4) kg, 1.75 (7.95) m] participated in two experimental sessions separated by one week. In these sessions, they performed three blocks of 10 putts trying to stop the golf ball at the center of a dartboard painted 2 m away. The performance was assessed depending on the area of the dartboard where the ball stopped, and the acceleration signals were acquired using the Xsens Dot. The results showed that to evaluate performance, 18 trials were necessary to reach reliable values using the 0–10 scoring system, and 28 trials were necessary for the 0–3 scoring system. Regarding the reliability of the accelerometer-related variables, 7 attempts were necessary to obtain good-to-excellent reliability values for most of the variables. It could be concluded that putting in medium-to-high handicap golf players can be reliably measured using the abovementioned protocol.