A late injury-time equaliser for Welwyn Garden City deprived Berkhamsted of their 100 per cent winning record in the heat on Bank Holiday Monday.
Joint bosses Steve Bateman and Lee Bircham made four changes to the side that progressed in the FA Cup 48 hours earlier.
Berko had plenty of chances in the first-half but somehow it remained goalless.
Hosts City played better in the second period but it was Berko who took the lead in the 70th minute when an incisive pass from Cameron Brooks put Lewis Toomey in and his right-wing cross was met at the back post by Jonathan Lacey who finished with a tidy shot.
More chances came and went for Berko and that profligacy came back to haunt The Comrades when three minutes into injury time City put everyone forward for a late corner.
Centre-back Carl Mensah got up well to head the leveller and leave Berko disappointed.
Despite not gaining maximum points, Berko are still in second spot in the early Southern League Division One Central table.
On Saturday Berko won their FA Cup preliminary round game 3-2 away at Wembley FC.
Berko opened the scoring when a Toomey lobbed pass split the centre backs for Lacey to round his man.
Alex Taylor then slid a ball to Lacey and his cross from the left was netted by Toomey to make it 2-0.
Wembley were thrown a lifeline when Josh Chamberlain gave a penalty away which was converted to make it 2-1 at half-time.
On the hour Toomey beat the offside trap and ran on to make it 3-1.
He later won a penalty after being brought down but put it over the bar.
Sub Yasmine Femi-Gill pulled the score back to 3-2 with a decent shot to set up a needlessly nervy finish but Berko saw the tie out.
The draw on Tuesday put Berko away at Southern League Premier Division Central side St Ives Town in the first qualifying round.
This Saturday Berko host Kidlington, before visiting Chesham United’s ground to play Aylesbury United next Wednesday.