Final answer:
Applying excessive pressure to the vein is most likely to allow reflux to occur during venipuncture.
Step-by-step explanation:
In venipuncture, reflux is the backward flow of blood. To prevent reflux, it is important to secure proper needle placement and blood flow. Among the options given, the most likely factor that would allow reflux to occur during venipuncture is 2) Applying excessive pressure to the vein. When excessive pressure is applied, it can cause the blood to flow backward, leading to reflux.