Current mayor Elaine O'Neal dropped the news in June that she would not seek re-election.
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Middleton was immediately pegged as a possible replacement, due to his current status on the Durham City Council.
However, in a social media post Wednesday, he announced that after "prayerful consideration" he decided not to throw his hat in the ring.
The candidate filing period closes at noon Friday. The primary election will happen on Oct. 10. The top two vote getters in the primary will then face off in the general election on Nov. 7.
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One of Middleton's fellow councilmembers DeDreana Freeman recently announced that she would be in the race.
In addition, North Carolina State Senator Mike Woodard, who served on the Durham City Council from 2005-2012 and has been in the state Senate since 2013, announced he would run for the office.
Durham passes city budget, with pay increase for firefighters, no hike in property taxes