Comparison of different DNA extraction methods for the molecular study of bark beetles (Coleoptera: Curculionidae, Scolytinae)

Abstract

Bark beetles are one of the most important pests in forests. Because of their small size and highly similar morphological characters, molecular approaches may be quite useful for a valid species determination. In this context molecular identification represents an accurate and modern method for species identification. The purity and high quantity of extracted DNA have important role in successful amplification of the target fragment of the genome. The aim of this study was comparing different DNA extraction methods in order to choose the highest quality and quantity of DNA extract for the identification of bark beetles. During the study bark beetles were collected from different parts of the North forests of Iran. Five different DNA extraction methods were performed and evaluated on individual specimen including Chelex, Phenol chloroform, CTAB, salting out and Lysis buffer in the laboratory. The quantity and quality of extracted DNA were measured by spectrophotometer and gel electrophoresis. The result of DNA quantity mean ranged between (23.6-579.7 ng/µl) and the mean quality which was measured by 260/280 ratio (0.9-1.8). The statistical analysis was done by SPSS software, revealing significant differences between extraction methods. The results suggested that Chelex and salting out showed the highest quantity of all used methods.