It was announced in May 2008 that Lisa de Cazotte would serve as Executive producer for season two, joined by Head writer Sri Rao. The 14 new episodes of Night Shift began taping in high-definition in June 2008, with the series airing Tuesdays at 11 p.m. and premiering on July 22, 2008. SOAPnet said the second season "will feature new and returning characters as well as the return of 'legacy' characters from GH. In addition, the continuity between story lines on Night Shift and GH will match."  The second season finished its run on October 21, 2008.
Prior to its premiere, SOAPnet had announced that Night Shift would "delve deeper into the relationships, friendships and medical cases seen at the hospital." It was noted that unlike General Hospital itself, the stories on Night Shift would be "self-contained and wrap up during each weekly one-hour episode," as well as being "understandable to viewers who do not watch General Hospital." With the goal "to attract younger viewers to both SOAPnet and General Hospital," characters would be "plucked from General Hospital's history" and "be mostly comprised of younger characters with ties to GH's core families."  Night Shift's storylines, however, did not directly intersect with those on General Hospital.