Passive stretching is commonly used to increase limb range of movement prior to athletic performance but it is unclear which component of the muscle-tendon unit (MTU) is affected by this procedure. Movement of the myotendinous junction (MTJ) of the gastrocnemius medialis muscle was measured by ultrasonography in eight male participants (20.5 ± 0.9 years) during a standard stretch in which the ankle was passively dorsiflexed at 1°.s^-1 from 0° (the foot at right angles to the tibia) to the participants’ volitional end range of motion (ROM). Passive torque and muscle fascicle length and pennation angle were also measured. Standard stretch measurements were made before (pre-) and after (post-) five passive conditioning stretches. During each conditioning stretch the MTU was taken to the end ROM and held for one minute. Pre-conditioning the extension of the MTU during stretch was taken up almost equally by muscle and tendon. Following conditioning ROM increased by 4.6 ± 1.5° (17%) and the passive stiffness of the MTU was reduced (between 20-25°) by 47% from 16.0 ± 3.6 to 10.2 ± 2.0 Nm.°^-1. Distal MTJ displacement (between 0-25°) increased from 0.92 ± 0.06 to 1.16 ± 0.05 cm, accounting for all the additional MTU elongation and indicating that there was no change in tendon properties. Muscle extension pre-conditioning was explicable by change in length and pennation angle of the fascicles but post-conditioning this was not the case suggesting that at least part of the change in muscle with conditioning stretches was due to altered properties of connective tissue.