The six-wheeled vehicle was built to operate just for three months. But it kept going and going until it was finally doomed by a ferocious dust storm eight months ago.
Flight controllers made numerous attempts to contact it and sent one final series of recovery commands on Tuesday night, accompanied by one last wake-up song, Billie Holiday's "I'll Be Seeing You."
There was no response, only silence.
Remarkably spry until communication ceased last June, Opportunity roamed a record 28 miles (45 kilometers) around Mars.
Opportunity and its long-dead twin Rover, Spirit, found evidence that ancient Mars had water flowing on its surface and might have been capable of sustaining microbial life.
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(With inputs from APTN)