Generation of Dna Barcode Data Base for Gunther's Loach Nemacheilus Guentheri (day, 1867)

By : Jaculine Pereira. J

Page No: 1-12

Abstract
The Gunther's loach, Nemacheilus guentheri (Day, 1867) was collected from the streams of Harangi river, Kalur village, Madikeri District of Karnataka State. Morphometric and meristic characterization was carried out for N. guentheri. The DNA was extracted from N. guentheri with both phenol chloroform method and HiPurA multi-Sample DNA purification kit. Purity of DNA was checked with UV spectrophotometer (260 nm and 280nm) and DNA was confirmed with Agarose gel Electrophoresis (1%). DNA barcoding was carried out for the DNA extracts using Cytochrome c Oxidase subunit 1(COI) gene. Polymerase Chain Reaction (PCR) was carried with COI gene primer and the PCR product size of 652bp was confirmed with Agarose gel electrophoresis (2%) using 100bp DNA ladder and DNA content was analyzed with Biophotometer (Eppendorf). The purity and concentration of DNA was determined at 260nm and 280 nm, the purity and concentration recorded for the PCR product were 1.58-1.63 (DNA quality is good) and 7.12 (4.6-10.1) respectively. PCR products of COI gene were sequenced and submitted to Genbank (NCBI), the accession numbers were provided by GenBank for N. guentheri includes KP772692, KP772693, KP772694 and KP772695.

Authors :
Jaculine Pereira. J, R. Jeya Shakila, B. Ahilan, K. Karal Marx, C.B.T. Rajagopalsamy and A. Ancy Jenifer
Fisheries College and Research Institute, Thoothukudi-628 008 Tamil Nadu Fisheries University, Tamil Nadu.