title: A simplified arthropod genomic-DNA extraction protocol for polymerase chain reaction (PCR)-based specimen identification through barcoding.
abstract: Genomic DNA extraction protocols generally require the use of expensive and hazardous reagents necessary for decontamination of phenolic compounds from the extracts. In addition, they are lengthy, hindering large-scale sample extractions necessary for high-throughput analyses. Here we describe a simple, time and cost-efficient method for genomic DNA extraction from insects. The extracted DNA was successfully used in a Polymerase Chain Reaction (PCR), making it suitable for automation for large-scale genetic analysis and barcoding studies. The protocol employs a single purification step to remove polysaccharides and other contaminating compounds using a non-hazardous reagent buffer. In addition, we conducted a bioinformatics database analysis as proof of concept for the efficiency of the DNA extraction protocol by using universal barcoding primers specific for cytochrome c oxidase I gene to identify different arthropod specimens through Barcode of Life Database (BOLD) database search. The usefulness of this protocol in various molecular biology and biodiversity studies is further discussed.
authors.affiliation: Department of Entomology, Purdue University, Smith Hall, 901 West State Street, West Lafayette, IN 47907, USA.
authors.forename: Venu M
authors.initials: VM
authors.lastname: Margam
authors.forename: Emma W
authors.initials: EW
authors.lastname: Gachomo
authors.forename: John H
authors.initials: JH
authors.lastname: Shukle
authors.forename: Oluwole O
authors.initials: OO
authors.lastname: Ariyo
authors.forename: Manfredo J
authors.initials: MJ
authors.lastname: Seufferheld
authors.forename: Simeon O
authors.initials: SO
authors.lastname: Kotchoni
mesh_terms: D000818:Animals; D001181:Arthropods / Q000145:classification* / Q000235:genetics*; D004247:DNA / Q000235:genetics* / Q000302:isolation & purification*; D058893:DNA Barcoding, Taxonomic / Q000379:methods*; D016678:Genome / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*
chemical_list: D004247:DNA