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 26th, 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 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.
In the men's 100m, Kishane Thompson scorched to a personal best of 9.82sec.
A year ago Thompson ran 9.91 seconds in the first round but did not show up for the semi-finals.
He tied Oblique Seville's personal best set earlier this month and the Jamaicans are joint second in the world behind Ferdinand Omanyala's world lead 9.79 seconds.
Defending champion Seville was his usual smooth self as he clocked 9.98 seconds while World Indoor 60m bronze medalist Ackeem Blake was the other runner under 10 seconds with 9.95 seconds (1.0m/s).
