Background: The examination of the lip print is known as Cheiloscopy. This is the method of identification of a person based on characteristic arrangement of lines appearing on the red part of the lips. Aim and Objective: This study was aimed at investigating gender variation in lip print pattern distribution of identical and non-identical twins. Methods: the study comprised 80 individuals (40 pairs- 25 pairs of non-identical and 15pairs of identical twins) of the Yoruba tribe of Nigeria (ranging between of ages of 5- 66 years) residing in Igbo-Ora community of Oyo state in Western Nigeria. The lips of the subjects were properly cleaned and a red lipstick was applied on it. After which, the lip print impression was made on a white plane sheet. Cellophane tape was then struck on it for permanent record. The obtained lip print was studied using a magnifying lens. Results and Discussion: The following were the results in identical twins, type I males (16.7%), females (12.5%); type I’ males and females 8.3%, type II males 33.3%, females 30.5%; type III males 18.8%, females 20.8%; type IV males I6.7%, females 18.1%; type V males and females 8.3%. whereas for the non-identical twins, type I males (18.8%), females (18.4%); type I’ males 7.8%, females 15.4%, type II males 31.2%, females 24.3%; type III males 21.9%, females 23.5%; type IV males I2.5%, females 14.0%; type V males 7.8% and females 4.4%. Conclusion: It is suggestive that certain pattern types have affinity for a particular gender probably due to hormonal interplay which ultimately results in variations in their distribution.