Comparison of different DNA extraction methods in bark beetles (Col.: Curculionidae) identification
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عنوان دوره: سومین کنگره بین المللی حشره شناسی ایران
نویسندگان
Department of Plant Protection, College of Agriculture and Natural Resources, University of Tehran, Karaj, Iran,
چکیده
Bark beetles belong to subfamily Scolytinae are one of the most important pest group in the forest worldwide. Because of similarity in morphological characters, and small sizes, molecular method can confirm bark beetle identification in species level. In molecular identification, extraction and purification of the DNA has an important role in amplification of target segment of different species. This study was done to find out the best method for obtaining the highest quality and quantity of DNA of bark beetles. Sampling was done by direct collection of insects from different parts of North forest of Iran. The sample moved to the tube 1.5 containing 96% alcohol and was taken to the laboratory. We evaluated four different DNA extraction methods including: Chelex, Phenol chloroform, CTAB and Salting out which all of them were set up according to published data. The quality and quantity of extracted DNA samples were measured by Spectrophotometer by 260 / 280. To ensure about the quality of extracted DNA gel electrophoresis has done by running much amount of DNA. The results of DNA quantity were calculated as 163.2, 140.5, 158.2 and 122.5 for Chelex, Sating out, CTAB and phenol chloroform respectively. The quality ranges were obtained as 1.5-1.8 (Chelex), 1.02-1.5 (Salting out), 1.5-1.3 (CTAB) and 1.01-1.2 (Phenol chloroform), respectively. The statistical analysis was done by SPSS software and showed that there are significant differences among the methods. The results suggested that Chelex and Salting out methods make the highest quantity and quality DNA from bark beetles
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