The palmaris longus is harvested as a tendon graft in various surgical procedures. Several tests are used to assess the presence of palmaris longus tendon. In the present study, we attempted to assess the interobserver and intraobserver reliability of five of the most famous methods and also the examination of fifth superficial flexor function. Two observers, who had been trained on the tests and had practiced them, examined 105 volunteers on two separate occasions and in 1-month interval the results were recorded. The reliability of each method was assessed with Kappa measurement. Kappa ranged from 0.541 (moderate reliability) to 0.813 (almost complete agreement) for palmaris. The highest interobserver and intraobserver reliability and also the best agreement with other tests were of Schaeffer. The lowest kappa was for Thompson and the others have good to excellent reliability. Kappa for interobserver and intraobserver reliability for the fifth flexor were 0.415 and 0.500 (moderate reliability), respectively. The tests that were assessed have good reliability except for Thompson that has a moderate one. It seems that the standard test (Schaeffer) is the best method for the assessment of the absence or presence of palmaris longus. The method used for the evaluation of fifth superficial flexor variations assessment has a moderate interobserver and intraobserver reliability.