Fraser-Pryce, crowned in Beijing in 2008 and in London in 2012, won her race in Kingston in 10.98sec (0.7 m/s).
At 37, the eight-time Olympic medalist is aiming for a new title at the Paris Olympics, which begin on July 26, and is gaining momentum after only resuming competition in mid-June.
Jackson, 29, a double world runner-up in the distance, was similarly comfortable in her her heat, winning in 10.99 (0.9 m/s).
The two sprinters were part of a resounding Jamaican hat-trick in Tokyo in 2021 behind Elaine Thompson-Herah, also crowned in Rio in 2016, who will not compete in the Paris Olympics after withdrawing from the trials on Wednesday due to injury.
Fraser-Pryce and Jackson meet on Friday for the semi-finals and the final, where they must take one of the first three places.
Over 400m, the surprise world champion from Budapest, 22-year-old Antonio Watson, pulled up after about 120m and will not compete in the Olympics.
