DNA barcoding is used to identify particular specie by Cytochrome c oxidase 1 (CO1) mitochondrial DNA sequence. It is also deal with genetic variability between species and its polymorphism. Principally phylogenetic methods based on maximum likelihood and neighbourhood joining tree for variability in sequence and taxonomical classification of species. DNA barcoding is the one means of establishing an effective and precise system for species identification. A standard short target of genetic marker often used to craft sequence contour of identified against unidentified species. Specified marker/tag of mitochondrial cytochrome C oxidase subunit 1 is used for the species identification purpose. Usefulness of following method lay off the divergence degree among species. Compare to previously used identification method through external or visible morphology, barcoding is more reliable. Computation of genetic distances and construction of Neighbor Joining tree based on the Kimura 2 Parameter method were also done systematically. Analysis of evolutionary tree stood clearly by making phylogenetic tree with relationships among their species, alike species bunching under the same tree node while differing species clustered under distinct nodes with no exemptions. Study findings concluded that sequence of Cytochrome C oxidase 1 used as a milestone to provide identification of species at molecular level, its similarity and divergence among the related species.