Broodiness is a behavioural trait which is observed in most common breeds of domestic and indigenous fowl. The onset and regulation of incubation behaviour is caused by this trait and due to this fundamental role in avian production, it has been of great interest to scientists, producers and breeders. The chicken Prolactin (PRL) gene has been accepted to have crucial effect in egg production as its increase in secretion causes the onset of incubation (broodiness). Therefore, the present study has been aimed to investigate the Single Nucleotide Polymorphisms (SNPs) in the PRL gene of native chicken ‘Zo-ar’ of Mizoram. DNA extraction followed by PCR-RFLP has been done in 50 blood samples collected from different parts of the state. Two alleles C and T have been detected for the PRL gene in the population. The T allele was found to be predominant in contrast to the C allele with a frequency of 0.66 and 0.34 respectively. The genotype frequencies of CC, CT and TT were found to be 0.06, 0.56 and 0.38 respectively in the studied population. The population conforming to the Hardy-Weinberg equilibrium indicates that no selection pressure was applied for any economic traits to the population in the past.