Acoustic myography (AMG) is a non-invasive method to assess muscle function during daily activities. AMG has great scope for assessment of musculoskeletal problems. The aim of this study was to create an AMG data set for general clinical use and relate these findings to age and gender. 10 healthy subjects (5 men/5 women), in each decade from 20 to 69 years of age (n = 50), were assessed. Their clinical health was tested. AMG measurements were carried out on muscles involved in defined movements of the upper and lower extremities. Muscle performance was measured using efficiency (E-score) and fibre recruitment (temporal (T-score) and spatial (S-score) summation). AMG-measurements showed good reproducibility. In each age group, it was found that for all those daily living skills measured, there was no gender difference. A walking and stair climbing test revealed that both legs are used equally and in a balanced way in healthy subjects. Moreover, there was no change in this function with increasing age up to 69 years. However, a cycling test with loading revealed that in elderly subjects the coordination of muscle use is impaired compared to that of the younger adults. Finally, a flexion test of the arm revealed an age-related decrease in the efficiency/coordination of m.Biceps alone, and a keyboard writing test suggests no effect on m.Trapezius. This reference data set now illustrates the reproducibility and ease of use of acoustic myography in the clinic and provides a means of assessing individuals with musculoskeletal problems.