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Dartford have turned to Tony Burman following the departures of Adam Flanagan and Jamie Coyle.
Burman master-minded Dartford's rise in non-league football since they moved to Princes Park and spent 14 years in charge.
He stepped down in May 2018 but has remained a club director and also works with Dartford's academy.
Flanagan and Coyle left the club following Tuesday night's 3-2 defeat at Tonbridge.
In a short statement issued by Dartford, the club also confirmed two further departures.
The club said: "Following the departure of Jamie Coyle and Adam Flanagan last night, we are also able to confirm that Goalkeeping Coach Andy Young and Strength and Conditioning Coach Harrison Grant have now left the club.
"We thank them all for their hard work and commitment to Dartford FC since joining and wish them all well in the future.
"We are pleased to announce that Tony Burman will take interim charge of all first team matters until a new permanent manager is appointed.
"The club is already receiving expressions of interest from suitably qualified candidates and a formal recruitment process will begin in due course."
Dartford are at home to Dorking Wanderers in National League South on Saturday. They have taken just seven points from their first nine games.