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Substitute Greg Wilkinson struck with his first touch to break Guiseley's hearts in the last minute of extra-time.
As penalties beckoned, Wilkinson steered the loose ball home after keeper Steve Drench had parried Kyle Perry's shot.
Altrincham had struck first in the 51st minute through James Lawrie. But the Lions hit back when Danny Forrest appeared to get the final touch to Jake Lawlor's header.
But the fantastic charge - which had seen Guiseley lose only once in 20 games - ended in dramatic fashion right at the death.
It's all over and Guiseley are beaten at the death. Altrincham fans flood the pitch as the visitors reflect on coming so close
That is heart-breaking for the visitors
Nightmare for Guiseley. Drench blocks from Perry as sub slips through but WILKINSON is following up to score with his first touch!
Two minutes of added time
Altrincham make a very late change as Wilkinson replaces Reeves. Obviously more confident of him taking a penalty ...
Perry curls shot on turn well wide. 30 seconds ....
Hows yer bottle @GuiseleyAFC @altrinchamfc ?— @brakes_updates 10 May 2014
Lawlor gets on the end of it but cannot direct the header this time
Parker cross is deflected, causing Coburn a real problem, and ball scrambled behind for a Guiseley corner. Two and a half minutes left...
Wilkinson was stripped and ready to take his place but the midfielder opts to continue
Richman down for Altrincham, maybe cramp as much as anything.
Ten minutes of extra-time to find a winner or it's down to penalties...
Boyes back defending to whack away then Drench fouled to end any pressure
Parker back in his own box with important clearance from Densmore cross. Corner to Alty
Parker makes surge towards edge of the box and plays ball into Boyes. He shoots on the turn but Coburn saves low to his left
The home crowd wake up with a roar and much stamping of feet as we begin part four...
It's half-time of extra-time...
Bower goes leaping down the touchline to protest after Potts is pulled back by the collar..
Altrincham making their second change, Tom Marshall comes on for left back Adam Griffin
Perry coming into his own in extra-time...
Perry rattles the bar from 30 yards! Guiseley breathe again...
A few tired legs and minds out there. The pitch is getting heavier
Holsgrove cross tipped nervously over the bar by Coburn. But Guiseley's corner comes to nothing
Ref has found his card now and can't wait to use it. Holdsworth become the fourth booking of the game for kicking the ball away
We're off once again and the rain seems to have stopped just in time
And that's it. We're heading for an extra 30 minutes!
We're two minutes into the three added on....
And now it's an Alty centre half in the book as Havern follows for dragging back Boyes
Ellis booked for foul on Clee just outside Guiseley box. Drench makes good save from Densmore free-kick
A sniff of a second for Guiseley. But Potts only managed a weak shot at Coburn after working his way cleverly into the box
Pity to see Forrest going off. Made a real impact in his half an hour
Guiseley double sub: Meynell for Forrest - must have picked up a knock and Holsgrove replaces Oli Johnson.
Forrest cuts in on his right foot and drills in skimming shot that Coburn has to concentrate with
Game getting scrappier. Eight minutes left and nobody wants to make a mistake
A big early chance for Perry as ball breaks to him unmarked in box but Drench makes crucial save...
Come on @GuiseleyAFC get a winner #Lions— @whitford369 10 May 2014
Walshaw subbed by Alty - moral victory for Guiseley there. Replaced by Kyle Perry
Guiseley pressing again and Coburn fails to hold Boyes cross
Guiseley level from the corner! Lawlor heads on and FORREST bundles home in the melee
Ball breaks to Forrest in the box. Brings it down promisingly and tries to place shot in bottom corner but deflection takes it wide for a corner
It looks like Guiseley have switched more to a 4-3-3 with Boyes as the spearhead up front and Forrest and Johnson either side.
Come on @GuiseleyAFC, turn it round— @alexhami17 10 May 2014
Shot from Clee clears the away end and disappears into someone's garden...
Comical corner conceded by Guiseley as Hall's tackle on Reeves pings off Ellis and flies behind
First change from Guiseley as Bower sends on Danny Forrest for Alex Johnson
Potts in the ref's book for flattening Clee in midfield...
A flare is lit in the Altrincham end but it fizzles out pretty quickly...
Clee plays ball into Walshaw who pulls it back for LAWRIE to steer past Drench from close range.
Good goal but might be stretching it to say it's "international class" as Altrincham radio claimed ....
1-0 to Altrincham! LAWRIE
Great chance again for Guiseley as Holdsworth free-kick flicked on to Ellis whose side-foot volley is beaten out by Coburn.
Teams are back on the pitch. Got extra-time written all over this...
Today's crowd is 4,632 - much higher than predicted. There are 794 away fans - six off what I said earlier....
Have destroyed a small toy because of the tension at the @GuiseleyAFC game. Lots of barking in the 2nd half. Hope they can hear me.— @MyloGuiseley 10 May 2014
Altrincham had the best of the first 20 minutes but Guiseley really came into it as the first half wore on.
Oli Johnson looking a real threat on the right wing and giving left back Adam Griffin a massive headache.
Referee John Brooks brings the half to an end, 0-0
Rothery bends cross into Altrincham goal mouth but it eludes everyone
One added minute
Slip by Ellis, who has been excellent up to now, but central defensive partner Hall is on hand to bale out his partner in the Guiseley box
Guiseley so close as Oli Johnson cuts past Griffin again into box and his close-range shot clips off keeper Coburn's leg and somehow avoids going in...
The game starting to open up now with five minutes left of first half
Great tackle from Ellis to intercept Warshaw flick-on as Reeves burst into the Guiseley box
Guiseley angry that Alex Johnson not getting any protection from some strong marking by Havern
Home keeper Coburn almost embarrassed by Rothery pot-shot from 30 yards and needs two grabs at it.
Altrincham captain Densmore shoots just over after being picked out in the box by Lawrie.
@GuiseleyAFC a view of the guiseley fans from the wrong end. http://t.co/ePdhfflwXZ— @Si_LUFC 10 May 2014
Walshaw unleashes an ambitious effort from way out but Drench gets everything safely behind it to save
Potts goes in with both feet on Clee. Referee John Brooks takes no more action than awarding free-kick
Oli Johnson eludes Griffin's tackle - and pull - to cut back to Alex Johnson who lifts shot over the bar. Best chance so far for Bower's men...
Some better possession football from Guiseley but Altrincham have men back in numbers
Another corner and Alty centre-half Leather jumps highest but heads well over
Hall, under pressure from Reeves, has trouble dealing with Moult's high ball and concedes a corner - which Ellis nods out of harm's way
Nice piece of skill by Alty left winger Clee but his cross eludes both Walshaw and Reeves.
Holdsworth's cross causes some confusion in Alty penalty area as centre half Havern heads ball out of his keeper's hands.
Then a wind-assisted long ball catches out Guiseley for another home corner
.... but not much fun for the Guiseley fans packed on the open terrace
Absolutely lashing down now. Rain in Manchester, eh, fancy that...
Reeves panics Drench into conceding a cheap throw-in near the Guiseley corner flag
Thumping effort by Jake Moult flies just past the post from 25 yards. Drench looked to have it covered
And so's this as Clee kick met firmly by the head of Potts at the near post.
Corner number three for the hosts. Previous two both defended comfortably
Now Alty have another one. Very even so far with one big chance squandered
Second Guiseley corner but it comes to nothing
Mark Bower looking very calm on the touchline ... for now
First opportunity for the visitors as Boyes volley deflected behind for a corner - which is headed over by Alty old boy Danny Hall
Big chance for Alty as Clee plays Reeves through on goal. But the former Farsley striker clears the bar with only Guiseley keeper Drench to beat
Lions skipper Andy Holdsworth gets told to calm it down by the ref after a late tackle on Adam Griffin
Hope my cousin @dannyellis555 and @GuiseleyAFC smash it today in Manchester.. #guiseley— @ScottRawnsley 10 May 2014
The ground looks absolutely rammed. People standing in the main stand and no sign of any gaps on the terraces
Danny Ellis heads away any danger
Very early corner for the home side....
Guiseley get us underway
Here they come..... Guiseley wearing all yellow today
Best of luck @GuiseleyAFC. Here's to realising your dream of playing Conference football and playing Stoke Reserves!— @SkyeDyson 10 May 2014
The nice shiny trophy has been put on its plinth - the only time you'll see that word in a match blog...
We're waiting for the teams now
Good atmosphere building here, plenty of singing from both sets of fans.
Good luck to @GuiseleyAFC today! #pushforpromotion— @AmyIlling10 10 May 2014
Fans still pouring in. It's rammed in the main stand and filling up on the other three sides.
Travelling Lions @GuiseleyAFC http://t.co/KZCbe5IC4D— @Feldm3ister 10 May 2014
Altrincham are unchanged from the side that scored two late goals here to topple Hednesford last week.
Guiseley must find a way of shackling old boy James Walshaw who has netted 34 times this season, including a hat-trick against his former club in the league meeting here in October.
ALTRINCHAM: Coburn, Densmore, Havern, Leather, Griffin, Lawrie, Moult, Richman, Clee, Walshaw, Reeves. Subs: Marshall, Doughty, Wilkinson, Perry, Saunders.
Best of luck to @GuiseleyAFC at @altrinchamfc today. It will be a tough game against Lee Sinnott's men but should be a cracking game.— @whiting_ian 10 May 2014
Guiseley are without two of their mainstays this afternoon.
Winger Wayne Brooksby, who scored last week's winner in the semi-final second leg at North Ferriby, misses out with a broken jaw. It will be the first game he hasn't played all season.
And player-coach Danny Boshell is barred from appearing as part of the agreement in the deal that took him back to Guiseley from Altrincham.
In their place come Oli Johnson and Michael Potts.
The third change sees Steve Drench return in goal in place of veteran Steve Dickinson after recovering from his virus.
GUISELEY: Drench, Holdsworth, Hall, Ellis, Parker, Lawlor, Potts, O Johnson, Boyes, A Johnson, Rothery. Subs: Meynell, Forrest, Harrison, Holsgrove, Dickinson.
Reckon the crowd will be around 3,000 today with over 800 from Guiseley. There were ten coaches laid on for free by club owner Steve Parkin.
RT @NavChohan1: Best of luck @GuiseleyAFC in the play off final today at Altrincham from @Shipley_College— @Shipley_College 10 May 2014
RT @altrinchamfc: The stage is set #Alty v @GuiseleyAFC. #SkrillNorth promotion final. http://t.co/p7e0kNMZiK— @RballBall 10 May 2014
Welcome from Moss Lane for the Skrill North play-off final as Guiseley bid for a place in the Conference.
Mark Bower's side have come through on an incredible run with only one defeat in their last 20 games. Now Lee Sinnott's Altrincham stand in their way.
After three successive play-off near misses, can the Lions make it fourth time lucky this afternoon?