Among 25 ties taking place, nine non-league sides, who had already negotiated the qualifying rounds, met clubs from Leagues One and Two (third and fourth tiers) for a place in the second round.
And with a history for shocks and stories of underdogs triumphing against adversity, it was Chester and Dover that provided the upsets.
Goals from Ben Heneghan and Craig Mahon gave Chester a 2-1 win away at League Two promotion-chasers Southend United while Stefan Payne was the hero for Dover when his first-half goal was enough for a 1-0 home victory over Morecambe.
Elsewhere, fourth-tier Cheltenham Town thrashed third tier Swindon Town 5-0 at home while 1912 FA Cup winners Barnsley of the third tier beat fourth tier Burton Albion by the same score to provide the stand-out results amongst the league teams.
League Two side Cambridge United beat Fleetwood Town 1-0 to dump out a side one league above them.
Brothers Elliot and Tom Bradbrook both scored as Dartford won 4-3 away at Bromley in a clash of the non-league teams, while in another seven-goal thriller Milton Keynes Dons, who beat Manchester United 4-0 in the Capital One (League) Cup in August, won 4-3 at League One rivals Port Vale.
Capital One Cup quarter-finalists Sheffield United struggled to transfer their cup form to the FA Cup and drew 0-0 away at fellow third-tier Crewe Alexandra, who are currently bottom of their league, to ensure a replay.
The first round kicked off on Friday when Warrington Town, the lowest-ranked team left in the competition and in the first round for the first time, beat Exeter City, 100 league places above them 1-0.
Twelve more ties take place on Sunday and Monday including 1987 winners Coventry City, now of League One, facing non-League Worcester City, with the draw for the next round taking place on Monday.
(Reporting By Sam Holden)