Southern segment

I-35 enters the state in Freeborn County south of Albert Lea. The first one mile (1.6 km) northbound contains a rest area and Minnesota welcome center. Shortly after the first exit, for County Road 5 (CR 5), there is a sweeping S-curve to the northwest and back to the north. The exit for CR 13 marks the last exit before Albert Lea. Up to here, the Interstate roughly marked the halfway point between U.S. Route 65 (US 65) and US 69. Both U.S. Highways end in Albert Lea.

In Albert Lea, it curves to the northeast and meets US 65, which passes beneath the Interstate. US 65 picks up Business Loop Interstate 35 (BL I-35), which follows the US Highway through the city. It then crosses Albert Lea Lake of a pair of 0.25-mile-long (0.40 km) bridges. It straightens back to the north to meet CR 46, which follows the former route of US 16. Shortly after that interchange, it intersects the national northern end of US 65; access to US 65 there is limited to the southbound lanes of I-35, and traffic from US 65 can only access the northbound lanes of I-35. At the northeastern edge of Albert Lea, there is an interchange with I-90, which directs traffic toward La Crosse, Wisconsin, or Sioux Falls, South Dakota.

I-35 continues north through the rolling hills of southern Minnesota. At Clarks Grove, it intersects MN 251. Between there and the CR 35 exit to Geneva, it passes to the west of Geneva Lake. Just east of Ellendale is an interchange with MN 30. Exit 32, CR 4 to Hope, is the last interchange before Owatonna. On the southern edge of Owatonna is a cloverleaf interchange with US 14. North of CR 45, the CR 34 exit serves northern Owatonna and provides access to the Owatonna Degner Regional Airport. The CR 9 exit serves Clinton Falls and a Cabela's store.

Further north, I-35 meets CR 12 and CR 23 in Medford. Along CR 23, which serves as a western frontage road to I-35, lies an outlet mall. Now in Rice County, the Interstate approaches Faribault. Exit 55 only serves exiting northbound traffic and entering southbound traffic. MN 60 is the main east–west street through Faribault, and MN 21 provides access to Faribault from the north of town. To the south, MN 21 ends at MN 60, but the road continues south and reconnects to I-35 at exit 55.

North of Faribault, the Interstate heads due north before the two directions are briefly separated when they enter a corridor of trees. The route straightens out again and intersects CR 1 and MN 19 at diamond interchanges spaced three miles (4.8 km) apart. It briefly enters Scott County where it meets CR 2 near Elko New Market. It then enters Dakota County and the Minneapolis–St. Paul metropolitan area.

In southwestern Lakeville, there is a folded diamond interchange with CR 70. Right before a partial cloverleaf interchange with CR 60, the Interstate crosses Marion Lake. Shortly after an interchange with CR 5 and CR 50, there is a park and ride complex on the northbound side of the Interstate. An entrance and exit ramp provide access to the Interstate while two entrances from Kendrick Avenue, a frontage road, provide non-Interstate access to the complex. After the CR 46 interchange, the Interstate enters Burnsville. A half diamond interchange, with only northbound exit and southbound entrance ramps, immediately precedes the split. I-35 splits into I-35W and I-35E (signed for Minneapolis and Saint Paul, respectively).

Northern segment

I-35W and I-35E join again at Columbus, near Forest Lake. The route then continues as I-35. In contrast to the rolling terrain of I-35 south of the Twin Cities area, this region is quite flat. This is the Anoka Sand Plain, an area that was formed by outwash under the last glaciers that lay over this area as they melted. This is an extensive region that continues north for another 20 miles (32 km). The urban sprawl of the northern Twin Cities continues along I-35 to North Branch.

In Pine City and Hinckley, there are museums near I-35: the Pine City one at a refurbished fur trading post and interpretive center and the Hinckley one commemorating the Great Hinckley Forest Fire that occurred in 1894. Motorists who take BL I-35 through Pine City can experience a quintessential all-American main street.

Between Sandstone and Moose Lake, the landscape around I-35 makes a dramatic change. Exposed granite can be seen at the surface, and the deciduous forest changes to evergreens.

I-35 climbs Thompson Hill as it approaches the city of Duluth. As the highway enters the city limits at Boundary Avenue, the Saint Louis Bay and Lake Superior are in view. I-35 then descends Thompson Hill into Duluth, with numerous exits. The freeway has an interchange with I-535/US 53, known locally as the "Can of Worms". The junction features a pair of left exits from I-35, a stoplight, and lane drops over the I-35 bridge.

I-35 then proceeds to downtown Duluth. After multiple exits, I-35 leaves the downtown area and follows the Lake Superior shoreline for two miles (3.2 km) to its national northern terminus at its junction with MN 61 and 26th Avenue East. At this junction where I-35 ends, MN 61 begins its route northeast along the North Shore of Lake Superior.

Transit

Jefferson Lines provides intercity bus service along the length of I-35 in Minnesota serving over a dozen communities along the route with transfer locations in Albert Lea, the Twin Cities, and Duluth.^[5]