Accurate data regarding the size of the erect penis is of great importance to several disciplines working with male patients, but little data exists on the best technique to measure penile length. While some previous small studies have suggested good correlation between stretched penile length, others have shown significant variability. Penile girth has been less well studied, and little data exist on the possible errors induced by differing observers and different techniques. Much of the published data report penile length measured from the penopubic skin junction-to-glans tip (STT) rather than pubic bone-to-tip (BTT). We wished to assess the accuracy of different techniques of penile measurements with multiple observers. Men who achieved full erection using dynamic penile Doppler ultrasound for the diagnosis of sexual dysfunction or a desire for objective penile measurement were included in the study. Exclusion criteria were penile scarring, curvature, or congenital abnormality. In each case, the penis was measured by one of the seven andrology specialists in a private air-conditioned (21 °C) environment. Each patient had three parameters measured: circumference (girth) of the penile shaft, length from suprapubic skin-to-distal glans (STT), and pubis-to-distal glans (BTT). The three measurements were recorded in the stretched flaccid state, and the same three measurements were then repeated in the fully erect state, following induction of full erection with intracavernosal injection. We analyzed the accuracy of each flaccid measurement using the erect measurements as a reference, for the overall patient population and for each observer. In total, 201 adult men (mean age 49.4 years) were included in this study. Assessing the penis in the stretched and flaccid state gave a mean underestimate of the erect measurement of ~20% (STT length 23.39%, BTT length 19.86%, and circumference 21.38%). In this large, multicenter, multi-observer study of penis size, flaccid measurements were only moderately accurate in predicting erect size. They were also significantly observer dependent. Measuring penile length from pubic bone to tip of glans is more accurate and reliable, the discrepancy being most notable in overweight patients.