The Kohathites, a branch of the tribe of Levi, held a significant role in the marching order of the Israelites as described in Numbers 10. Positioned after Reuben, Simeon, and Gad, the Kohathites were tasked with carrying the sacred furniture of the tabernacle. This included the burnt altar, the incense altar, the laver, the candelabra, the mercy seat, and the ark. Their responsibility was distinct from other Levite branches like Gershon and Merari, who carried the structural components of the tabernacle itself. The placement of the Kohathites and other Levites in the marching order ensured they were surrounded and protected by the other tribes, highlighting the importance of their duties in safeguarding the holy items during the journey through the wilderness.

The Kohathites, a specific group within the Levites, were assigned the sacred duty of carrying the Ark of the Covenant, the most revered object among the Israelites, representing the Throne of God. According to explicit instructions, the Ark was not to be transported on a cart or pulled by oxen but was to be carried with long poles thrust through rings built into the Ark, made only of acacia wood. The Kohathites were the only ones permitted to carry it, and they were forbidden even to look upon it. Aaron and the priests were to cover the Ark before the Kohathites could approach to transport it. In a notable incident recorded in I Chronicles 13, during the moving of the Ark under King David's direction, virtually everything was done contrary to these commands. As the oxen stumbled and the Ark appeared to be falling, Uzza reached out to steady it, an act that resulted in his immediate death by divine judgment. Whether Uzza was a Kohathite is not specified, but his action was deemed the final act of arrogance and presumptuousness, defiling the Ark by the touch of a sinning man, contrary to the holiness required in handling such a sacred object. This event underscores the strict standards of obedience and holiness imposed on those, like the Kohathites, who were tasked with drawing near to God in their service.

The Kohathites, as part of the Levitical tribe, held a significant responsibility concerning the most holy objects of Israelite worship. In the incident involving Uzza and the Ark of the Covenant, it is noted that the Kohathites were not even supposed to look at the uncovered Ark. Their strict instructions underscored the reverence required for such sacred items. When Uzza touched the Ark to prevent it from falling, despite not being indicated as a Kohathite, it was seen as the final act of desecration, arrogance, and presumption, leading to a swift divine judgment. This event illustrates God's strict requirement for conformity to His directives regarding holy objects, emphasizing the seriousness of presumption in handling what is sacred to Him.