Ayahuasca ceremonies are held in the ceremonial house at a specified time at night set by the master shaman. This is an arbitrary hour traditional to each lineage of Ayahuasca medicine, usually between 6:30 p.m. and 10:00 p.m. The ceremonies typically last three to six hours in duration but have been known to continue for up to ten hours. The ceremony is a time for spiritual transformation and spiritual growth, as the shaman practices the ancient medicine, guiding the ceremony through the use of icaros (sacred medicine songs).

Each participant will have a cushion on which to sit or lie down or a chair, as well as a pillow, blanket, vomit receptacle, roll of toilet paper to wipe the mouth, and cup of water for rinsing and spitting.

The ceremony begins as the master shaman or shamans sing icaros into the bottle of Ayahuasca to set the intent of the ceremony. This protects the apprentices, patients, participants, shamans, and their mesa and sets the sacred space for healing. The doctor spirits are called to awaken in the medicine.

Each individual cup is then poured one by one, and icaros in the name of the participant are sung into the cup. This protects them and sets the intent for their purification, learning, healing, and cleansing. The cup is then presented to the participant. This is a time for the participant to focus their intent and purpose for participating in the ceremony. This may be done through prayer or thought. The Ayahuasca is then drunk, and the cup is passed back to the shaman. The process is repeated for each remaining participant. The shamans drink last.

As the effects of the Ayahuasca take hold, the lights are extinguished and remain off until the end of the ceremony. The ceremonies are held in the dark because the eyes become very sensitive to light. It is typical to have the ability to see even though all participants sit in total darkness.

When the lights are turned off, the shamans begin to sing icaros and play shacapas, leaf rattles used to carry the visions of the ceremony and direct the medicine. The icaros are sung steadily for the first few hours of the ceremony and then are sung intermittently toward the end.  

The ceremony ends when the lights are turned on and the effects of the Ayahuasca have dropped considerably.