An Efficient Method for DNA Extraction from Preserved Stoneflies (Insecta, Plecoptera)

Abstract

Recent studies of ecology and taxonomy on stoneflies employed molecular sequences data based on DNA amplification used both field and museum alcohol-preserved specimens. In this study, we compared four different DNA extraction protocols based on preserved stoneflies specimens with SDS-Ethanol method, Phenol/Cloroform, Phenol/Chloroform/Isoamyl Alcohol method and modification of this last method to obtain best efficiency DNA extraction method. The absence of PCR inhibitors and the DNA quality were evaluated by PCR amplification of the mitochondrial 12S DNA gene using insects-specific primers. DNA extraction efficiency using method 4 is significantly higher with the highest quality >2.19 (260/280 nm) and quantity (>1370 ng/μl) and significantly difference (P< 0.05) than DNA extraction efficiency using other methods. This technique provides alternative DNA extraction methods for preserved stoneflies.