Room 314 is an episodic story about five different, unrelated couples who occupy the same room of a hotel, each facing challenges in their relationship. In one episode, two co-workers, Jack (director Michael Knowles) and Kathy (Robyn Myhr), contemplate having an affair but are racked with guilt. Another involves Nick (Matthew Del Negro), a man about to be married, and Stacey (Joelle Carter), a woman he picked up at his bachelor party. David and Caly (Michael Mosley and Jennifer Marlowe) are two former lovers attending a friend's wedding, and Matt and Tracey (Todd Swenson and Monique Vukovic) are a couple who may be a hooker and her client, although the film doesn't explain. Harry (Michael Laurence) is suicidal and his wife Gretchen (Sarah Jenkins) tries to find out why. Each story deals with some aspect of sexual relationships and their difficulties. In addition to acting in and directing the film, Knowles also wrote Room 314 and served as the cinematographer.