The Vagina Monologues is made up of a varying number of monologues read by a varying number of women (initially, Eve Ensler performed every monologue herself, with subsequent performances featuring three actresses, and more recent versions featuring a different actress for every role). Every monologue somehow relates to the vagina, be it through sex, love, rape, menstruation, mutilation, masturbation, birth, orgasm, the variety of names for the vagina, or simply as a physical aspect of the body. A recurring theme throughout the piece is the vagina as a tool of female empowerment, and the ultimate embodiment of individuality. Some monologues include:

• I Was Twelve, My Mother Slapped Me: a chorus describing many young women's and girls' first menstrual period.
• My Angry Vagina, in which a woman humorously rants about injustices wrought against the vagina, such as tampons, douches, and the tools used by OB/GYNs
• My Vagina Was My Village, a monologue compiled from the testimonies of Bosnian women subjected to rape camps.
• The Little Coochie Snorcher That Could, in which a woman recalls memories of traumatic sexual experiences in her childhood and a self-described "positive healing" sexual experience in her adolescent years with an older woman. In the original version, she is 13, but later versions would change her age to 16. This particular skit has sparked numerous controversies and criticisms due to its content (see below).
• Reclaiming Cunt, a piece narrated by a woman who illustrates that the word "cunt" itself is a lovely word despite its disconcerting connotations
• The Woman Who Loved to Make Vaginas Happy, in which a sex worker for women discusses the intriguing details of her career and her love of giving women pleasure. In several performances it often comes at the end of the play, literally climaxing with a vocal demonstration of a "triple orgasm."
• Because He Liked to Look At It, in which a woman describes how she had thought her pubic area was ugly and had been embarrassed to even think about it, but changed her mind because of a sexual experience with a man named Bob who liked to spend hours looking at it.
• I Was There In The Room, a monologue in which Eve Ensler describes the birth of her granddaughter.

Every year a new monologue is added to highlight a current issue affecting women around the world. The monologue is performed at thousands of local V-Day benefit productions of the play that take place annually in February and March raising funds for local groups, shelters, crisis centers working to end violence against women. In 2003, for example, Ensler wrote a new monologue about the plight of women in Afghanistan under Taliban rule. This Monologue is known as "Under the Burqa."

All proceeds benefit the Sexual Assault Crisis & Support Center & the Women of the democratic Republic of Congo.