Enhanced DNA Extraction Protocol for Larvae and Adult Specimens of Clostera cupreata (Butler) (Lepidoptera: Notodontidae): A Key Leaf Defoliator of Poplar Trees

The CTAB DNA isolation protocol for both larval and adult specimens of Clostera cupreata has optimized with modifications included adjusting the concentrations of EDTA, SDS, and NaCl; substituting liquid nitrogen with extraction buffer for sample homogenization; and extending the incubation, mixing, and precipitation time period. This modified protocol ease of use and efficiency allow for the quick extraction of high-quality, high-yield entire genomic DNA using common reagents and equipment used in molecular laboratories. This will enable the documentation of genetic variability based on biotypes, facilitating studies on the emergence, distribution, and population dynamics of C. cupreata. To preserve the integrity of the extracted DNA, stringent measures are implemented to prevent contamination from various biological by-products, including proteins, salts, phenols, and polysaccharides. We have verified the quality and purity of the obtained DNA samples through standard quality assessments. The A260/A280 ratios consistently fall within the range of 1.80 to 1.88, providing further validation of the integrity and suitability of the isolated DNA by performing polymerase chain reaction analysis using mitochondrial COI primer. This approach is suitable for a wide range of molecular applications. Our extraction protocol yields high-quality DNA that may be used for sophisticated molecular processes like gene cloning, conventional polymerase chain reaction for genotyping, next-generation sequencing and barcode synthesis. Furthermore, this approach has potential as a preventive diagnostic tool for detecting invasive lepidopteran larval stages.