title: Comparative analysis of protocols for DNA extraction from soybean caterpillars.
abstract: Genomic DNA extraction is crucial for molecular research, including diagnostic and genome characterization of different organisms. The aim of this study was to comparatively analyze protocols of DNA extraction based on cell lysis by sarcosyl, cetyltrimethylammonium bromide, and sodium dodecyl sulfate, and to determine the most efficient method applicable to soybean caterpillars. DNA was extracted from specimens of Chrysodeixis includens and Spodoptera eridania using the aforementioned three methods. DNA quantification was performed using spectrophotometry and high molecular weight DNA ladders. The purity of the extracted DNA was determined by calculating the A260/A280 ratio. Cost and time for each DNA extraction method were estimated and analyzed statistically. The amount of DNA extracted by these three methods was sufficient for PCR amplification. The sarcosyl method yielded DNA of higher purity, because it generated a clearer pellet without viscosity, and yielded high quality amplification products of the COI gene I. The sarcosyl method showed lower cost per extraction and did not differ from the other methods with respect to preparation times. Cell lysis by sarcosyl represents the best method for DNA extraction in terms of yield, quality, and cost effectiveness.
authors.affiliation: Setor de Entomologia, Cooperativa Central Gaúcha Ltda, Unidade de Tecnologia, Cruz Alta, RS, Brasil.
authors.forename: J
authors.initials: J
authors.lastname: Palma
authors.affiliation: Centro de Ciências Rurais, Universidade Federal de Santa Maria, Santa Maria, RS, Brasil.
authors.forename: I
authors.initials: I
authors.lastname: Valmorbida
authors.affiliation: Programa de Pós-Graduação em Agronomia, Centro de Ciências Rurais, Clínica Fitossanitária, Departamento de Defesa Fitossanitária, Universidade Federal de Santa Maria, Santa Maria, RS, Brasil.
authors.forename: I F D
authors.initials: IF
authors.lastname: da Costa
authors.affiliation: Programa de Pós-Graduação em Agronomia, Centro de Ciências Rurais, Laboratório de Manejo Integrado de Pragas, Departamento de Defesa Fitossanitária, Universidade Federal de Santa Maria, Santa Maria, RS, Brasil.
authors.forename: J V C
authors.initials: JV
authors.lastname: Guedes
mesh_terms: D000818:Animals; D002458:Cell Fractionation / Q000379:methods*; D000077286:Cetrimonium; D002593:Cetrimonium Compounds / Q000737:chemistry; D004247:DNA / Q000737:chemistry* / Q000235:genetics; D017344:Genes, Insect; D016133:Polymerase Chain Reaction / Q000379:methods; D012967:Sodium Dodecyl Sulfate / Q000737:chemistry; D018411:Spodoptera / Q000737:chemistry* / Q000235:genetics
chemical_list: D002593:Cetrimonium Compounds; D012967:Sodium Dodecyl Sulfate; D004247:DNA; D000077286:Cetrimonium