Prior to moving to New York City, Lynch lived in San Francisco, California, where she performed at the drag club Trannyshack and reached the finals of the Miss Trannyshack pageant. It was her experience at Trannyshack that inspired her to start the club Knockoff on the Lower East Side of Manhattan, intended as a "knockoff" version of Trannyshack. She joined Scissor Sisters, founded by Jake Shears and Babydaddy, after they played their first live gig at the club.
She has stated that in her eyes, the objective of the band is "about people displaying their fantasies on the outside, trying to break out of the everyday, and look like their dreams." In the fan community for the Scissor Sisters, those fans who hold a special affinity for Ana are known as "nuns". A unique following of both homosexual men and heterosexual women who are attracted to Ana term themselves "Anasexual".
In 2005, she was featured on the New Order single, "Jetstream". The single was taken from the band's album, Waiting for the Sirens' Call. In 2009, Matronic performed on the glass harmonicaat the New Museum in NYC in Auroville, a multimedia ritual tribute to an experimental community dedicated to the guru Sri Aurobindo. Ana appears as a guest vocalist on the Mark Ronsonproduced track 'Safe (In the Heat of the Moment)' from the December 21, 2010, release All You Need Is Now from the band Duran Duran.
It was announced on 4 April 2012 by the BBC that Ana would appear as an advisor on the new UK version of The Voice, pairing up to assist Jessie J. On 7 March 2013, it was announced that Matronic would replace Sara Cox as commentator of the Eurovision Song Contest 2013 semi-finals in Malmö, Sweden for the United Kingdom.
In 2004, she revealed to British magazine Glamour that her parents' marriage broke up when she was a toddler due to her father's homosexuality, and that her father died of an AIDS-related illness when she was 15. She also stated in this interview that her immersion in gay culture is probably an attempt to feel closer to him, and understand who her father was as a person.
Matronic married her long-time boyfriend Seth Kirby in April 2010, at New York City Hall after having been together for seven years. Her stage name is said to be due to "a deep and abiding love of robots". She has a large tattoo on her right shoulder of bionic circuitry. She has a second tattoo on her ankle of a Celtic knot to signify her Irish roots.