Doncaster secured at least a play-off place and stayed top of League One after a hard-fought victory courtesy of Rob Jones' eighth goal of the season.
The Doncaster skipper fired home after latching onto Jamie McCombe's header from a David Cotterill free-kick.
Jean-Louis Akpa Akpro had Tranmere's best chance, heading Andy Robinson's free-kick wide late in the first-half.
The visitors had most of the play after going behind, but failed to create a clear-cut chance and stayed seventh.
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Doncaster manager Brian Flynn told BBC Radio Sheffield:
"This was going to be a key week and we've come out with six points, clean sheets in both games and a thoroughly good performance.
"We were in control, but we didn't create many chances.
"It was a good second half and Rob [Jones] finished well - that's two centre-halves scoring winning goals this week."