On September 6, 1988, Daphne Ashbrook with her then husband, Lorenzo Lamas, gave birth to their daughter Paton Lee Lamas. Her current partner is Director and Producer David Vito Gregoli.

A 2004 trip to the United Kingdom was filmed for the 2005 documentary Daphne Ashbrook in the UK. This DVD documentary followed her work with Doctor Who, including her role as Perfection in the Big Finish Productions audio play The Next Life. She performed opposite Paul McGann for first time since the 1996 telemovie. In 2006, she played Charlotte Howell in the audio drama Dark Shadows: The Book of Temptation for the same company. She has been to the UK twice thus far for Doctor Who convention events. She was featured in extensive interviews on the Doctor Who podcast Doctor Who: DWO Whocast and Doctor Who podcast Doctor Who: Podshock. Additionally, she featured in other podcasts including "The Happiness Patrol".

Known for both her comic timing and deep emotional portrayals, she has appeared in many television and film roles including Star Trek: Deep Space Nine (as the titular character in "Melora") and the Doctor Who telemovie, in which she played the role of Doctor Grace Holloway. Her portrayal as Grace in Doctor Who caused consternation among some of that show's fans because she was the first character to romantically kiss the Doctor, setting a precedent for the more sexually charged nature of the new Doctor Who series that began in 2005.

Her past TV and film work includes Cold Case, CSI, Crossing Jordan, JAG, Murder She Wrote, Judging Amy, acclaimed mini series Intruders with Richard Crenna, Guardian, a recurring role on The O.C. and a lead role in the police drama Family Honor with Ray Liotta and Eli Wallach. Her recent roles include a part in David Ondaatje's 2009 motion picture The Lodger starring Alfred Molina, Shane West, Simon Baker and Hope Davis, and guest starring roles in NCIS reuniting with Charlie Grace star Mark Harmon, Ghost Whisperer, with Jennifer Love Hewitt, and Without a Trace with Anthony LaPaglia and Steven Weber. She guest starred as a Dancer visiting from (the former) Czechoslovakia in the third season of Fame. She is currently (2012) starring in the new Nickelodeon night time soap Hollywood Heights as Chloe Carter (Melissa Ordway) mom, Jackie.

In 2010 she released her first music CD, Grace Notes on Dharmapala Records to very positive reviews. The songs range from Roots to Jazz and are mostly covers from some of her favorite singer/songwriters, such as Joni Mitchell and Patty Griffin. The CD was produced by composer/multi-instrumentalist David Vito Gregoli. She has performed music from the CD at various conventions and other events.

In 2012 Ashbrook released a memoir Dead Woman Laughing (An actor's take from both sides of the camera). It details her life as an actor and her experience growing up in an acting family. The book has received very good reviews.