Rodent behavioral test results (open field, anxiety, and cognition assays) are notoriously irreproducible across labs and even across cohorts in the same lab due to uncontrolled confounds.

Control the following variables to reduce behavioral variability: 1) Test during the active (dark) phase, which is more translationally relevant than light-phase testing. 2) Avoid testing on cage-change days; the disruption of a cage change measurably affects behavior for hours afterward. 3) Handle mice via acrylic tube or cupped-hand methods instead of tail-lifting; tail-handled mice showed reduced exploration of novel objects in testing. 4) Keep room humidity in the 40-60% range and standardize/document light levels used during testing. 5) Keep the same handler (and handler sex) consistent across a cohort, and document test order and inter-test spacing when running a battery of assays.