Present investigation was conducted to measure reliable information about genetic differences between male and female individuals. RAPD technique was used to access the genetic diversity using 12 oligo decamer primer generating 127 loci (average 10.5 loci per oligo primer). Number of polymorphic bands found was 106. Average genetic diversity was 52.5 percent, ranged from 16 (between WCh1 and WPR4) to 91 percent (between WLg3 and WLg4). Mean band presence per individual was 51.68±23.0. PIC value ranged from 0.30 (OPT-20) to 0.44 (OPM-13) banding pattern 3 to 19 types with primer discriminatory power ranged between 0.35 to 0.99. SHAN Cluster produced three major cluster, first cluster of male with eight accession, second cluster of female with WPR and RIR accession, and third cluster with four accessions of WLg groups. Present investigation demonstrated that application of RAPD to distinguish the male and female bird’s accessions and two different female accessions too. Hence RAPD may be utilized for identification of male and female chicken spices.