The Glovers couldn't make it three consecutive wins as Lincoln City left Somerset with all three points in Sky Bet League Two.
After an even start at Huish Park the Imps took the lead with a little over 15 minutes of the half remaining when Danny Rowe converted from close range.
Having more than been in the game until the opener, Lincoln's strike did little to deter the hosts who continued to try and continue their run of improved form but on the stroke of half-time the deficit was doubled.
Rowe was the right man in the right place once again to collect a ball in the box before turning and lashing past Nathan Baxter to give the visitors some breathing space at the break.
After the restart, the hosts struggled to gather momentum against a well-drilled Lincoln outfit and were unable to make it three wins on the bounce, falling to defeat as the visitors extended their lead at the top.
Coming into the midweek encounter off the back of consecutive victories, Darren Way made two changes for the visit of the Danny Cowley's side.
Requiring stitches to a head wound on Saturday, defender Adel Gafaiti dropped from the squad to allow Bevis Mugabi a starting berth, whilst Yoann Arquin replaced Rhys Browne on the wing in a tactical change.
Two Academy products in the form of Gabby Rogers and Daniel Ojo made the bench, along with Matt Worthington who awaited his first appearance since rejoining the club last Friday.
On a bitterly cold night in Somerset it took both sides a bit of time to warm-up to the occasion, with the opening 10 minutes passing with little action of note.
John Akinde and Rowe both had tame efforts comfortably gathered by Nathan Baxter whilst at the other end Gary Warren glanced a teasing Alex Dobre cross wide of the target.
Needing a spark to bring the game to life, it almost came courtesy of the infectious running of Sessi D'Almeida.
The Benin international charged into the Imps half after exchanging passes with Dobre, raced along the byline and forced Grant Smith to save with his legs from an acute angle.
More than matching their top of the league opponents the Glovers looked comfortable heading towards the half-hour mark, but were dealt a blow on 27 minutes.
Having been held at arm's length the visitors found a way past a resolute Town defence as Neal Eardley's cross from the right was perfect for Rowe to turn home at the near post to open the scoring.
Although ahead, the goal did little to alter the pattern of the match with both sides seeing most of their possession in the middle of the park.
The Glovers kept plugging away in search of an equaliser as Bournemouth loanee Dobre looked the most threatening in an advanced position, but chances remained at a premium for both sets of players.
However, with Cowley's side looking comfortable in possession one moment of precision saw the Imps make it 2-0 in first-half stoppage time.
Michael O'Connor spotted a free Rowe in the box and his clever through ball allowed the attacker to swivel and fire home from 12 yards for his second of the match with virtually the last play of the half.
Forced into a change at the break, Worthington was introduced for his first appearance since signing for the club on a permanent basis in place of Gary Warren, in a swap that saw Tom James move back into defence.
Once the game resumed the opening 10 minutes of the half belonged to Lincoln, with Harry Anderson close to scoring but for a reaction save from Baxter.
Needing a response at the other end the green and whites almost got one thanks to a solo effort.
The lively Dobre cut in from the left wing and engineered space for a venomous shot which whizzed just over the bar as the hosts registered their first attack of the second 45.
Requiring a big final half-an-hour top scorer Alex Fisher took the place of Francois Zoko up front in a bid to set up a dramatic final act.
Eager to get back into the match Town continued to battle and swung a number of balls into the box, but against a firm Lincoln defence there was nothing to show for it in front of goal.
At the other end and with an array of attacking talent, the Imps continued to threaten in spells, with Anderson firing just wide from range before Shay McCartan saw a shot cleared off the line by Jake Gray following a corner.
Academy product Gabby Rogers made his second EFL appearance for the club from the bench inside the final 10 minutes to add a positive note to proceedings, but there was to be no way back for the Glovers who suffered a first defeat in three.
Yeovil Town: Baxter; Grant, Warren (Worthington 46'), Mugabi, Dickinson; D'Almeida, James, Gray (Rogers 84'); Arquin, Zoko (Fisher 60'), Dobre.
Bookings: Warren 41', James 62', Arquin 90'
Subs not used: Nelson, Ojo, Santos, Browne.
Lincoln City: Smith; Eardley, Bolger, Shackell, Toffolo; Anderson, Bostwick, O'Connor (Frecklington 63'), Pett; Rowe (McCartan 67'), Akinde (Rhead 71').
Bookings: Frecklington 80', Toffolo 86'
Subs not used: Vickers, Chapman, Wilson, Brown.