Optimisation of DNA isolation and PCR techniques for beetle (Order: Coleoptera) specimens

DNA isolation is critical in many molecular biology studies, including genetics and genomics. Various DNA isolation techniques have been established to extract DNA from beetle species with DNA isolation kits available on the market. The polymerase chain reaction (PCR) success rate depends on the quality of DNA extracted from biological samples. In this study, four DNA isolation methods were compared, Chelex 100, N-cetyl-N, N, N,—trimethyl ammonium bromide (CTAB), Sodium Dodecyl Sulphate (SDS), and Tris, NaCl, EDTA, SDS (TNES), to determine the amount and quality of DNA obtained, the amount of time it took, and the effects of ethanol volume, incubation time, and temperature on DNA precipitation. The Chelex 100 method was the most successful compared to other methods and proved to be the superior DNA isolation method. We obtained the best amplification results for the beetle cytochrome oxidase 1 (CO1) gene at A1 Master Mix concentrations and A5 PCR cycle conditions. This study provides a road map for selecting the optimum DNA isolation methods for beetle species based on DNA quality, quantity, time consumption, cost, protein contamination, and success rate. The DNA isolated by the four DNA isolation techniques was suitable for further molecular studies like conservation genetics and genomics.