We identified a site suitable for venipuncture on the tail of the Virginia opossum (Didelphis virginiana) that is reliably and easily located. The prominent hemal arch associated with the ventral surface of caudal vertebra 5 serves as an easily palpated anatomical landmark for locating the ventral caudal vein for blood collection. Because this venipuncture site is only thinly covered by fur and visualization of the vein is not necessary for its location, site preparation and total animal handling time for routine venipuncture are minimal. Blood may be collected from immature and adult male and female animals, and the technique is easily taught to new technicians with minimal danger of injury to the animal.