The showdown between triple Olympic champion Bolt and world champion Blake lived up to its billing as the highlight of the four-day meeting as 'The Beast' roared home in a dazzling 9.75 seconds, a personal best and fastest in the world this year.
The time also made Blake the fourth fastest man of all time behind Bolt (9.58), American Tyson Gay (9.69) and Asafa Powell (9.72).
"No pressure at all... everything is good. I'm just fortunate," Blake said after eclipsing his personal best of 9.82. "I'm the national champion of Jamaica now, I go into the Olympics like this."
Bolt, who was unable to reel in his training partner, settled for second in 9.86 followed by former world record holder Powell in 9.88.
All three men qualified for the Jamaican Olympic team and will line up against a formidable American contingent at the London Games starting on July 27.
There will be no rest for Bolt and Blake, however, who will return to the track on Saturday looking for a spot in the 200m.
"We are right where we want to be going into London," Glen Mills, who coaches both Blake and Bolt, told Reuters.
"We just want to keep them healthy. The strategy, we didn't send him (Blake) to Europe and he is in far better shape than Bolt at this time.
"We have four weeks (until the Olympics) and we will take it in stride. We know what to do, so we'll get there.
"Bolt is a tough cookie and I think he will survive."
The men were not the only ones delivering excitement as women's 100m Olympic champion Shelly-Ann Fraser-Pryce provided some magic of her own by blazing to victory in 10.70 seconds, equalling the seventh fastest time in the event.
Only Americans Florence Griffith-Joyner, Carmelita Jeter and Marion Jones have run faster.
Veronica Campbell-Brown, a double Olympic gold medalist in the 200m, was a distant second in 10.82 with Kerron Stewart third on 10.94.
"I set a national record tonight and there is nothing else I could ask for," Fraser-Pryce said.
"I'm looking forward to the Olympics and defending my championship. It has been in the works. My coach said I could break the record and I did it.
"I worked really hard for this. I am not afraid of anybody."
The Bolt-Blake showdown left a capacity crowd that included Prime Minister Portia Simpson-Miller on the edge of their seats with the high-speed drama building steadily throughout a calm, muggy evening.
The semi-finals got the vuvuzelas blowing and drums pounding before Fraser-Pryce set the stage for the evening finale with her electrifying run.
The race marked the first meeting between the training partners since last year's world championships when Blake claimed Bolt's crown after the latter was disqualified from the final for a false start.
There were no such gifts on offer on Friday but Blake showed the Daegu win was no fluke when he exploded from the blocks and could not be caught.
"I thought it was a good race but everybody kept moving in the blocks and that kind of threw me off," Bolt said.
"I think overall I executed through the first 60 metres but the last part not so good, overall it was OK.
"For me it is just one of those things."
As well as the sprinters, a Jamaican team that is expected to light up the London Olympics continued to take shape with 400m hurdles Olympic and world champion Melanie Walker securing her spot by winning in an impressive 54.77.