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We examined the electromyographic (EMG) activity of the supraspinatus and other rotator cuff muscles, the three portions of the deltoid muscle, and the pectoralis major muscle in two previously suggested positions for isolating the supraspinatus. The position suggested by Jobe and colleagues is with the elbow extended, the shoulder in full internal rotation, and the arm in the scapular plane. Blackburn and colleagues recommended the prone position, with the elbow extended and the arm abducted to 100° and externally rotated. Fine-wire EMG activity was obtained from the rotator cuff muscles and surface EMG from the other muscles in 17 subjects tested in these two positions. Both positions resulted in significant activity of the supraspinatus, but the difference between these two positions was not statistically significant. The Jobe position produced greater activation of the anterior deltoid and pectoralis major, whereas the Blackburn position caused greater activation of the posterior deltoid. Both positions produced significant activation of the middle deltoid. We conclude that either position can be used to strengthen the supraspinatus; however, neither position selectively isolates the supraspinatus during manual muscle testing.