From 19 August, Caroline Barker will host the programme from 0530-0600.
Since it was first aired by BBC London in 2006, it has examined the issues affecting non-league football.
The show will be available via the BBC iPlayer all week, and a longer version will be released via free download from the Radio 5 live website.
Barker said: "With more fans of our level of football than ever before, this is a fantastic opportunity to show off the work of the clubs and, most importantly, the fans that make those teams tick."
The Non League Show will provide those supporters from across the country a new platform to send in their views on the grassroots game.
You can listen to BBC Radio 5 live via digital radio, online at bbc.co.uk/5live, Digital Freeview, Sky, Freesat, Virgin Media and 909 & 693 AM.