The western terminus of the route is at an intersection with NY 90 near the northern village line of Union Springs.

Its eastern terminus is at a junction with U.S. Route 20 (US 20) and NY 5 just west of the Auburn city limits in the town of Aurelius.

It briefly parallels a former alignment of itself, now known as "Old Route 326", before exiting Union Springs and entering the surrounding town of Springport.

Much of Springport, a town situated on the eastern shore of Cayuga Lake, is rural and largely undeveloped.

[3] NY 326 was assigned as part of the 1930 renumbering of state highways in New York and extended from the village of Union Springs to the city of Auburn.

[5][6][7] By 1977, US 20 and NY 5 were realigned once again to follow a new arterial leading from the north end of the divided highway eastward into downtown.

[8] On April 1, 1981, NY 326 was realigned between the hamlets of Oakwood and Half Acre as the result of a highway maintenance swap between the state of New York and Cayuga County.