In the 30-second spot, a narrator speaks over a picture of the White House.
"No matter who the president is, New Hampshire needs a strong voice in the U.S. Senate," the narrator continues.
While the ad does not explicitly signal that the Chamber expects Trump to lose the election, it makes the possibility very clear to viewers, touting Ayotte's ability to work "across the aisle" regardless of which party ends up winning the presidency.
A Chamber source told CNN that the ad is set to run first in New Hampshire, but noted that it could be the first in a series to run in other states as well.
Watch the spot below: