Wrexham's hopes of a historic fourth consecutive promotion were ended with a 2-2 Sky Bet Championship draw against Middlesbrough at the SToK Cae Ras.
Hull took advantage of Wrexham - who were playing National League football three years ago - dropping points to secure sixth spot with a 2-1 win over Norwich.
Middlesbrough will head to the play-offs after finishing fifth, with Southampton awaiting them in the semi-final, while Wrexham end up in seventh.
On a see-saw afternoon in which they started inside the top six, Wrexham were quickly outside it when Tommy Conway put Boro ahead after four minutes.
Wrexham were back above the dotted line when Josh Windass and Sam Smith struck, but David Strelec equalised just before the end of an eventful first half.
Neither side could find the winner which would have impacted them - fourth place would have given Boro semi-final home advantage in the second leg - but it was not for the want of trying.
Wrexham's fast start, which saw Ollie Rathbone take aim from 30 yards and Boro pair Adilson Malanda and Sol Brynn involved in a potentially costly mix-up, evaporated after four minutes.
Morgan Whittaker sent Callum Brittain haring away and his cross was turned in by Conway from close range to leave questions directed at goalkeeper Danny Ward.
Celebrating Boro fans would not have welcomed news of Ipswich's early 2-0 lead that left their automatic promotion hopes a distant prospect, and they fell silent after 28 minutes.
Smith was fouled to present Windass with a shooting opportunity from nearly 30 yards out, and Wrexham's top scorer curled the ball away from Brynn and into the top left-hand corner for his 17th goal of the season in all competitions.
Whittaker headed over an Ayling centre as Boro regathered their senses, but the visitors were soon behind.
Issa Kabore raided down the right to deliver a cross that was met by a bullet header from Smith and Wrexham were back in the top six.
Two minutes later they were back outside as Strelec was left unmarked from Matt Targett's cross to score with a cushioned volley that left Ward helpless.
The second half began in open fashion with Brynn clutching efforts from Smith and Rathbone and Ayling trying his luck from distance at the other end.
Boro came close to regaining the lead from a well-worked move, with Strelec denied from close range first by Ward and then Callum Doyle on the goal line.
Wrexham sent on Kieffer Moore and Nathan Broadhead, with Hull leading and the Tigers now holding the play-off spot.
But Boro were also pushing and Ayling's header drifted inches wide of the far post.
In a thrilling finale Smith failed to make a clean connection from an inviting ball, Windass fired over from the edge of the box and Moore shot wildly as the Wrexham winner refused to come.
Play-off semi-final first legs
Play-off semi-final second legs
Play-off final
Wrexham's Phil Parkinson:
"We've taken the fight to the wire and it just wasn't to be on the day.
"I don't think there's much more we could have asked from the lads against such a good side.
"To respond the way we did after going a goal down was a great effort.
"We didn't want the season to end because we've enjoyed it so much. It has, but to finish seventh is a real achievement at this level."
Wrexham co-owner Ryan Reynolds on Instagram:
"I am completely gutted by today's result but incredibly proud of our season.
"We've come a long way in five years and this was the best result in our 150+ year history. More to do. But for now, we have so much to be proud of, Reds."
Middlesbrough's Kim Hellberg:
"We knew there was very little chance of getting in the automatics, but when you know that's not possible you are not too down.
"We are going into the play-offs and this is a great chance for us.
"Four teams, 25 per cent chance - a brilliant chance - but Southampton are a very good football team.
"We're disappointed we couldn't win the game, but lots of good things came from it."
