Comparative isolation and amplification of Cytochrome oxidase 1 DNA from Oncorhynchus mykiss (Rainbow Trout) of Azad Jammu & Kashmir

Abstract

DNA extraction plays a pivotal role in the polymerase chain reaction (PCR) based molecular and genetic characterization methods. In order to find out the exact genetic lineage of the trout fish found in AJK, the fish DNA was extracted and analyzed further. Due to complicatedness regarding the fish DNA extraction, three different extraction methods were employed. We extracted genomic DNA from Rainbow trout by using standard phenol Chloroform method, a modified phenol Chloroform method and DNAzol DNA extraction method. The yield and quality of DNA extracted through different methods was compared. It was found that the extraction made by treating the samples with sucrose during initial steps of DNA extraction (Sucrose, Tris HCl (pH 7.5), MgCl2, 1%V/V Triton, SDS and proteinase K) showed the best results while DNAzol method was the least productive. Likewise the modified method was also found to be the most productive in terms of PCR yield of cytochrome oxidase 1 gene.