Every minute of every day, a baby is born. Most babies are easily seen to be a girl or a boy. Imagine how confusing it must be when we don’t know the sex of a newborn? This is rare and it can be very upsetting for parents. During the present study, a total of 47 individuals with unassigned sex were enrolled for chromosome study. These 47 individuals in different age group were categorized into three groups namely ambiguous genitalia, Intersex and Hypospadias. In Ambiguous genitalia group, a total of seventeen individuals were enrolled for chromosome study. Of these, thirteen individuals reared as males were found to have 46, XY karyotype. In the remaining four cases reared as female sex, three were found to have 46, XX karyotype and one which was phenotypically a female was found to have 46, XY karyotype. In Intersex group, a total of fourteen individuals were enrolled for chromosome study. Of these fourteen cases of Intersex reared as male sex by their parents, thirteen were found to possess 46, XY karyotype where as one cases was found possessing 46, XX. In Hypospadias group, a total of sixteen individuals were enrolled for chromosome study and were found to have 46, XY karyotype. The purpose of the present study was to find out the chromosomal sex in these individuals. The study is first of its kind from Jammu region of Jammu and Kashmir, India.