DNA Barcoding of Cyprinid Fish Chagunius chagunio Hamilton,1822 from Phewa Lake, Nepal

Abstract

The present study is the first of its type that uses a technique of DNA barcoding to determine identification and relationship of a species of fish from Phewa lake, Nepal. The mitochondrial DNA from two ethanol-preserved samples of fish, randomly collected from Phewa lake, was extracted using Gene AllExgene TMtissue extraction kit. 650 base pair of mitochondrial cytochrome c oxidase subunit 1 (CO1) was amplified using a cocktail of four primers and was sequenced bidirectionaly using Sanger sequence method. The DNA sequences were edited using AliView software. The sequences confirmed Chagunius chagunio as their alignment with 16 reference sequences belonging to Chagunius chagunio in the NCBI GenBank, scored highest percentage of Query Cover (75% to 100%) and Percentage Identity (97.29% to 100%). The MEGA software analysed the DNA sequences to obtain their corresponding protein sequences. The DNA sequences were submitted to the GenBank and accession numbers (MN087472 and MN087473) were obtained. Clustal Omega software analysed multiple sequence alignment among 19 homologous DNA sequences of Chagunius chagunio from India, Bangladesh and Phewa lake, Nepal. The percentage of similarity among the aligned sequences was calculated as 39.3%. Based on the neighbour joining tree, the Chagunius chagunio of Phewa lake is found closely related with Chagunius chagunio of Bangladesh.