New York State Route 327 (NY 327) is a state highway in New York State. It begins at the NY 13/NY 34/NY 96 concurrency in the Town of Ithaca and ends at NY 79 in the Town of Enfield. It is located entirely within Tompkins County.

Route 327 begins at an intersection with Routes 13, 34, and 96 just outside the city of Ithaca. The highway is outside the urbanized city, and is rural and mountainous for most of its length. The route, after beginning, heads to the southwest as Enfield Falls Road. After parallelling the three-way concurrency, Route 327 turns west at an intersection with a short connector to Robert H. Treman State Park. The highway, about 1,000 feet (300 m) above sea level, runs along the park border to the south, and the higher mountain to the north.

Route 327 intersects with Trumbulls Corners Road and turns northward towards Bostwick Corners after crossing over Connecticut Hill. Just north of Bostwick Corners, Route 327 terminates at an intersection with NY 79 in Enifeld.

NY 327 was assigned as part of the 1930 renumbering of state highways in New York and has not been substantially altered since.

The entire route is in Tompkins County.