Filter paper for preservation, storage, and distribution of insect and pathogen DNA samples.
Proper DNA storage is critical for studies involving genetic analysis of insects and for molecular diagnostics of pathogens carried by them. Molecular surveillance of pathogens carried by insects can involve screening of thousands of insect DNA samples. Problems with storage and degradation of these samples can arise. In this study, a simple filter paper-based method for storage and preservation of insect DNA was evaluated using polymerase chain reaction (PCR). DNA was isolated from individual house fly, Musca domestica L., adults by using a cell lysis technique. From 50 house flies known to carry Campylobacter spp., a portion of the DNA sample was stored frozen and another portion was pipetted onto filter paper. At monthly intervals for 7 mo, PCR was conducted using 1 microl of the frozen DNA sample and a 2.0-mm disk from the filter paper samples as the PCR template. Two markers were used, a 450-bp region of the insect mitochondrial DNA (mtDNA) ND5 gene and a 857-bp region of the Campylobacter spp. mtDNA 16S rDNA gene. PCR amplification was successful for all of the samples regardless of the storage method. The filter paper method is a simple and economical way to store, preserve, and distribute DNA samples for PCR analysis.
Owens
Carrie B
CB
Department of Entomology, University of Arkansas, Fayetteville, AR 72701, USA.
Szalanski
Allen L
AL
D000818:Animals; D002167:Campylobacter / Q000235:genetics*; D004247:DNA / Q000032:analysis; D004269:DNA, Bacterial / Q000032:analysis; D004272:DNA, Mitochondrial / Q000032:analysis; D006793:Houseflies / Q000235:genetics*; D007303:Insect Vectors / Q000235:genetics; D010209:Paper; D016133:Polymerase Chain Reaction; D013048:Specimen Handling / Q000379:methods*
D004269:DNA, Bacterial; D004272:DNA, Mitochondrial; D004247:DNA