Mma Precious Ramotswe: Owner of the No. 1 Ladies' Detective Agency in Gaborone, Botswana. She is self-taught as a detective, and generally successful in resolving the cases brought to her. In her youth, she married a musician who was cruel to her. She had a baby who died soon after birth, and learned she could have no more children. She is in her thirties. She is engaged to marry Mr. J.L.B. Matekoni. She drives a tiny white van which she has had for 11 years.
Obed Ramotswe: Late father of Precious, who was a strong and wise man. She loves him dearly and often thinks of him as she solves her cases, and as she decides to become engaged to Mr. J.L.B. Matekoni and accept the orphans he took in. He left her a herd of 2,000 cattle; she sold most of them to start her business. She kept about 60, which a cousin watches for her.
Mr. J.L.B. Matekoni: Owner of Tlokweng Road Speedy Motors garage, and possibly the best mechanic in Botswana. He is a very kind man, in his forties. He suffers from depression when this novel opens.
Mma Grace Makutsi: She is the secretary to Mma Ramotswe, very efficient, known for scoring 97 on her final exam at the secretarial college. She wears large glasses. She is caring for one of her brothers, dying from disease. She is in her twenties. She has been given the title of assistant detective, and acting assistant manager at the garage.
Richard: Brother of Mma Makutsi, who now lives with her in her rented room on the Lobatse Road. He is very ill, with a terminal illness.
Mma Silvia Potokwane: Matron of the orphan farm outside Gaborone. She is a most persuasive woman, in seeking help for her orphans and keeping the place running. She finds a quiet place for Mr. J.L.B. Matekoni to recuperate.
Motholeli: Thirteen-year-old girl taken as foster child by Mr. J.L.B. Matekoni. Her mother died when she was 7. The custom of her people was to bury a newborn still nursing with the mother, if the mother died for any reason. Motholeli recovered her brother from the shallow grave and headed for the road, where a kind couple picked her up. She and the baby were in hospital, as Motholeli had tuberculosis and the boy needed care. Recovered from the worst of it, the two are taken home by a woman at the hospital. She has a recurrence of TB that leaves her unable to walk, and she uses a wheelchair. When that family must relocate for the husband's job, she and her young brother are placed at the orphan farm, and then taken as foster children by Mr. J.L.B. Matekoni. She writes her own story for school, saying she has had three lives.
Puso: Five years younger brother of Motholeli. He is a lively boy, well-behaved like his sister, who has been caring for him since his birth. He and his sister are of the Basarwa people of Gaborone, hunter-gatherers who live in the bush. Both are foster children to Mr. J.L.B. Matekoni, and cared for by Mma Ramotswe in this first month away from the orphan farm.
Rose: Maid to Mma Ramotswe. She is a conscientious and reliable woman who has worked for Mma Ramotswe since she bought the house on Zebra Drive. She aids in the care of the foster children, and has children of her own.
Big Government Man: He seeks the aid of the No. 1 Ladies Detective Agency, to learn who is poisoning his younger brother. His name is never stated, but his appearances in the newspapers and power in government are described.
Dr. Moffat: Diagnoses and treats Mr. J.L.B. Matekoni for depression, which he says is a common disease, and he has good medicines to treat him.
Family of the big government man: His father, mother, brother and sister-in-law live in the main family home on their cattle farm.
Samuel the cook: Man who sought work on the farm, but was assigned to be the cook instead, and who dislikes being cook, regardless of his talents.
Useless young apprentices: Two young men offered the opportunity to learn the art of car mechanic at Speedy Motors. Both are young, about 19, and their main interest is meeting girls in bars.
Mataila: Boy of 5 or 6 years, brought to the orphan farm wearing no clothes, having no words, not knowing his name. Mma Potokwane gave him a name, and tries to reach him. He bites, like a wild boy, and the first people who found him said he smelled like a lion. Mma Ramotswe meets him early on, and Mr. J.L.B. Matekoni connects with him toward the end of the novel.
Mr. Moemedi“Two Shots” Pulani: Businessman of Gaborone who loves to look at beautiful women, and runs beauty contests in Botswana. He seeks help from the Detective Agency, because one of the five finalists for Miss Beauty and Integrity has been caught stealing. Can Mma Makutsi learn more about the four remaining finalists, to assure him a true winner, with integrity?
Motlamedi: Finalist in the Miss Beauty and Integrity contest who is attending university, but is a "good time girl" known to the garage apprentice, and in Mma Makutsi's view, a "bad girl".
Patricia: Finalist in the Miss Beauty and Integrity contest who lives in Tlokweng with her mother. She hopes to attend the Botswana Secretarial College. Mma Makutsi judges her as having integrity.
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