Developing the DNA extraction method for seahorse

Abstract

DNA extraction is one of the most important steps in the process of DNA or gene analysis to date. Phenol- Chloroform and Salting out methods have been kept using in small or less modern- equipped laboratories in developing countries due to the low cost and the amount of DNA yield. In this study, DNA extraction was carried out in a species of seahorses named Hippocampus kuda which is the spotted seahorse and commonly grown in the central areas of Vietnam. Two DNA extraction methods were performed and compared to select the best method. The evaluation of extraction method was based on the DNA quality, DNA quantities. The quality of DNA was evaluated by the presence of DNA on agarose gel and the ability to amplify PCR products. A sequence of 247 base pairs of the mitochondrial (mt) DNA region was analyzed to test the ability to be amplified of extracted DNA by PCR. The study has also focused on the optimization of the techniques utilized in DNA extraction from tail tissue or fin tissue of live seahorses without scarifying them. The data collected is 30% success of Phenol- Chloroform method as compared with over 70% success of Salting out method. In addition, Salting out method requires working on less hazardous chemicals and lowering in term of cost and supplies. Therefore, the salt out method became the core to be optimized and improved; then, a complete technique can be withdrawn and announced. Keywords: Hippocampus kuda, Phenol: Chloroform method, salting out method, Mitochondrial (mt) DNA control region.